catalogo cell biolabs

Cell-Based Assays

Ordering Information & Support 4

5

Stem Cell Research 31

Viral Expression 39

microRNA Analysis 73

TRANSFORMATION ADHESION MIGRATION INVASION WOUND HEALING

2 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500

TABLE OF CONTENTS

CELL HEALTH CELL HYPOXIA PHAGOCYTOSIS CONTRACTION ANGIOGENESIS

IPS CELL REPROGRAMMING RETROVIRAL EXPRESSION SYSTEMS FEEDER CELLS COLONY FORMATION ASSAYS ALKALINE PHOSPHATASE ASSAYS

ADENO-ASSOCIATED VIRUS (AAV)

ADENOVIRUS LENTIVIRUS RETROVIRUS

PREMADE VIRUSES EXPRESSION SYSTEMS PURIFICATION KITS TITER KITS TRANSDUCTION KITS

PRECURSOR CLONE COLLECTION MAMMALIAN EXPRESSION VECTORS VIRAL EXPRESSION PLASMIDS FUNCTIONAL REPORTER SYSTEM KNOCKDOWN ENHANCER

3

Oxidative Stress / Damage 83

Cell Signaling & Protein Biology 111

Product Index

Worldwide Contacts

151

157

TABLE OF CONTENTS

www.cellbiolabs.com [email protected]

Metabolism Research 129

Pathogen and Toxin Assays 147

PROTEIN OXIDATION/NITRATION LIPID PEROXIDATION DNA/RNA DAMAGE AND REPAIR HYPOXIA ASSAYS REACTIVE OXYGEN SPECIES (ROS) ASSAYS ANTIOXIDANT ASSAYS

SMALL GTPASE / G-PROTEIN SIGNALING KINASE ASSAYS REPORTER CELLS AND REAGENTS EPITOPE TAGS PROTEIN ANALYSIS TOOLS

CHOLESTEROL/CHOLESTERYL ESTER ASSAYS APOLIPOPROTEIN ASSAYS AND REAGENTS FATTY ACID METABOLISM ASSAYS SERUM PROTEIN ASSAYS RENAL FUNCTION ASSAYS ALCOHOL ASSAYS

VIRUS CORE ANTIGEN ASSAYS TOXIN ASSAYS

ORDERING INFORMATION & SUPPORT

4

Worldwide Technical Support Placing an Order within the U.S.

Placing an Order outside the U.S.

Our Customer Service representatives are available Monday through Friday from 8 am to 5 pm Pacific Time. Most orders received by 2 pm Pacific Time will be shipped the same day. We will notify you immediately of any backordered item. We accept VISA®, MasterCard® and Ameri-can Express® cards. Net 30 day terms may be offered upon credit approval.

Phone 1 858 271 6500 1 888 CBL 0505 (Toll-Free) Fax 1 858 271 6514 E-mail [email protected] Online www.cellbiolabs.com Mail Attn: Customer Service 7758 Arjons Drive San Diego, CA 92126

Pricing

Kit Components

Do you have questions about a particular product before you buy? Do you need help with a protocol? Our Technical Service Scientists have been directly involved in the development and testing of our products, so you get the bene-fit of their hands-on experience.

Phone 1 858 271 6500 1 888 CBL 0505 (US Toll-Free) Fax 1 858 271 6514 E-mail [email protected]

Current U.S. prices are available online at www.cellbiolabs.com, or you may request a separate price list by e-mail. Prices are sub-ject to change without notice. For pricing outside the U.S. please contact your local distributor, which can be found on the inside back cover of this catalog.

Because our kits are QC tested by lot, indi-vidual kit components are generally not available for purchase separately. However, certain components may be available in bulk quantities on a custom basis. For more information please inquire by sending a message to [email protected].

Please see our Worldwide Distributors section on the inside back cover of this cata-log. We have a network of global distributors serving life science researchers in over 70 countries. If you don’t see your country listed, please contact our U.S. office.

Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500

6

CytoSelect™ 96-Well Cell Transformation Assay—Traditional Soft Agar Colony Formation

Product Name Detection Size Catalog Number

CytoSelect™ 96-Well Cell Transformation Assay (Soft Agar Colony Formation)

Fluorometric 1 Plate* CBA-130

5 Plates* CBA-130-5

Our CytoSelect™ 96-Well Cell Transformation Assay (Soft Agar Colony Formation) is suitable for measur-ing malignant transformation where no downstream analysis is required. Transformed cells cannot be re-covered; however, no manual cell counting is re-quired. With this assay, cells are incubated in a semisolid agar medium for 6-8 days, then solubilized, lysed and detected using CyQuant® GR dye in a fluorometric plate reader.

Fast Results: 6-8 days vs. 21 days Plate Reader Convenience: Eliminates manual

counting Versatile Format: Designed for 96-well through-

put, but can be adapted for 48, 24, 12 or 6-well

CELL-BASED ASSAYS Colony Formation Assays

Transformation of normal cells into neoplastic cells results in a population capable of pro-liferating independently of internal and external signals that normally restrain growth. The soft agar colony formation assay has traditionally been used to monitor anchorage-independent growth, employing 3-4 weeks of cell growth followed by manual cell counting.

We have advanced the soft agar assay to eliminate tedious manual cell counting, allow high-throughput drug screening, and enable recovery of transformed cells for downstream analysis. These advances have also allowed us to develop a unique kit for the separation of clonogenic cancer cells from normal cells in heterogeneous solid tumors.

Tumor Cell / Soft Agar Assays

Cell Transformation Assay Principle.

Recent Product Citations 1. Eisfeld, A. et al. (2014). NRAS isoforms differentially affect

downstream pathways, cell growth, and cell transformation. PNAS 111:4179-4184.

2. Gong, J. et al. (2014). Combined targeting of STAT3/NFkB/COX-2/EP4 for effective management of pancreatic cancer. Clin. Cancer Res. 20:1259-1273.

3. Gupta, P. et al. (2013). Ocrasin targets the JNK-NFkB axis to sensitize glioma cells to TNF-alpha-induced apoptosis. Car-cinogenesis 34:388-396.

4. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103.

5. Rai, V. et al. (2012). Lysophosphatidic acid targets vascular and oncogenic pathways via RAGE signaling. J. Exp. Med. 209:2339-2350.

6. Ghantous, A. et al. (2012). Inhibition of tumor promotion by parthenolide: epigenetic modulation of p21. Cancer Prev. Res. 5:1298-1309.

7. Dennis, M. et al. (2012). Snail controls the mesenchymal phe-notype and drives erlotinib resistance in oral epithelial and head and neck squamous cell carcinoma cells. Carcinogenesis 147:726-732.


*Each kit provides sufficient reagent quantities to perform 96, 48, 24, 12, or 6 tests in a 96, 48, 24, 12, or 6-well plate, respectively.

CytoSelect™ 96-Well Cell Transformation Assays—Advanced Soft Agar with Post-Incubation Cell Recovery The CytoSelect™ 96-Well Cell Transformation Assay (Cell Recovery Compatible) provides a robust system for screening oncogenes and cell transformation in-hibitors. Transformed cells may be recovered for fur-ther downstream analysis following colony formation.

Faster Results: 6-8 days vs. 21 days Cell Recovery: Transformed cells remain viable

for further analysis Plate Reader Convenience: Eliminates manual

counting of cells Versatile Format: Designed for 96-well through-

put, but can be adapted for 48, 24, 12 or 6-well

Cell Transformation Assay Principle. Cell colonies form after a 6-8 day incubation with agar matrix. Transformed cells can then be either lysed and detected with a fluorescent dye or recovered and re-plated.

*Each kit provides sufficient reagent quantities to perform 96, 48, 24, 12, or 6 tests in a 96, 48, 24, 12, or 6-well plate, respectively. **The 384-well kit does not allow for cell recovery due to small well size. ***Each kit provides sufficient reagents for one or five 384-well plates respectively.


CytoSelect™ 96-Well Cell Transformation Assay (Cell Recovery Compatible)

Colorimetric 1 Plate* CBA-135

5 Plates* CBA-135-5

Fluorometric 1 Plate* CBA-140

5 Plates* CBA-140-5

CytoSelect™ 384-Well Cell Transformation Assay** 1 Plate*** CBA-145

5 Plates*** CBA-145-5 Fluorometric

7


Recent Product Citations 1. Singh, R. et al. (2013). Increasing the complexity of chromatin:

functionally distinct roles for replication-dependent histone H2A isoforms in cell proliferation and carcinogenesis. Nucleic Acids Res. 10.1093/nar/gkt736. (CBA-135)

2. Shukla, A. et al. (2013). Extracellular signal-regulated kinase 5: a potential therapeutic target for malignant mesotheliomas. Clin. Cancer Res. 19:2071-2083. (CBA-135)

3. Niccoli, S. et al. (2012). The Asian-American E6 variant protein of human papillomavirus 16 alone is sufficient to promote im-mortalization, transformation, and migration of primary human foreskin keratinocytes. J. Virol. 86:12384-12396. (CBA-135)

4. Hong, S.W. et al. (2012). Ring finger protein 149 is an E3 ubiq-uitin ligase active on wild-type v-Raf murine sarcoma viral on-cogene homolog B1 (BRAF). J. Biol. Chem. 287:24017-24025. (CBA-135)

5. Lee, H.J. et al. (2012). Cheokine (C-X-C motif) ligand 12 is associated with gallbladder carcinoma progression and is a novel independent poor prognostic factor. Clin. Cancer Res. 18:3270-3280. (CBA-135)

6. Chapeau, E.A. et al. (2012). Ecotropic viral integration site 1 (EVI1) regulates multiple cellular processes important for can-cer and is a synergistic partner for FOS proteinin invasive tu-mors. PNAS 109:2168-2173. (CBA-135)

7. Lim, S.K. et al. (2011). Tyrosine phosphorylation of transcrip-tional coactivator WW-domain binding protein 2 regulates es-trogen receptor alpha function in breast cancer via the Wnt pathway. FASEB J. 25:3004-3018. (CBA-135)

8. Mathew, B. et al. (2011). The novel role of the mu opioid recep-tor in lung cancer progression: A laboratory investigation. Anesth. Analg. 112:558-567. (CBA-135)

9. Hirata, H. et al. (2010). Role of secreted Frizzled-related pro-tein3 in human renal cell carcinoma. Cancer Res. 70:1896-1905. (CBA-135)

10. Hirata, H. et al. (2009). Wnt antagonist gene DKK2 is epigen-etically silenced and inhibits renal cancer progression through apoptotic and cell cycle pathways. Clin. Cancer Res. 15:5678-5687. (CBA-135)



CytoSelect™ 96-Well In Vitro Tumor Sensitivity Assay Colorimetric 96 Assays CBA-150

5 x 96 Assays CBA-150-5

CytoSelect™ 96-Well In Vitro Tumor Sensitivity Assay

Inhibition of HeLa Cell Anchorage-Independent Growth by Taxol. HeLa cells were cultured for 7 days in the absence (top) or presence (bottom) of 1 nM Taxol according to the assay protocol.

The CytoSelect™ In Vitro Tumor Sensitivity Assay provides a stringent, anchorage-independent model for chemosensitivity testing and possible anticancer drug screening. The assay uses a soft agar matrix to promote the colony formation of neoplastic cells in about a week. Cells are quantified using a standard ELISA plate reader.

Fast Results: 6-8 days In Vivo Simulation: Resembles a three-

dimensional cell environment Plate Reader Convenience: Eliminates manual

counting

Tumor Sensitivity Assay Principle.

8


Recent Product Citations 1. Bard-Chapeau, E. et al. (2013). EVI1 oncoprotein interacts with

a large and complex network of proteins and integrates signals through protein phosphorylation. PNAS 110:E2885-E2894.

2. Takezawa, K. et al. (2012). HER2 amplification: a potential mechanism of acquired resistance to EGFR inhibition in EGFR-mutant lung cancers that lack the second-site EGFRT790M mutation. Cancer Discovery 2:922-933.

3. Li,C. et al. (2012). The root bark of Paeonia moutan is a poten-tial anticancer agent in human oral squamous cell carcinoma cells. Anticancer Res. 32:2625-2630.

4. Itamochi, H. et al. (2011). Inhibiting the mTOR pathway syner-gistically enhances cytotoxicity in ovarian cancer cells induced by etoposide through upregulation of c-Jun. Clin. Cancer Res. 17:4742-4750.

5. Kang, D.W. et al. (2010). Phospholipase D1 drives a positive feedback loop to reinforce the Wnt/ß-catenin/TCF signaling axis. Cancer Res. 70:4233-4242.


CytoSelect™ Clonogenic Tumor Cell Isolation Kit

9

Product Name Size Catalog Number

CytoSelect™ Clonogenic Tumor Cell Isolation Kit 5 Preps CBA-155

25 Preps CBA-155-5

Clonogenic Colony Formation, Isolation and Re-plating. A: Clonogenic colony formation (red arrows) and single cells (black arrows) after 7 day incubation. B: Isolation of clonogenic colonies from single cells. C: Re-plated clonogenic colonies after 3 days (no trypsinization). D: Re-plated clonogenic colonies 1 day after trypsinization.

Clean separation of clonogenic tumor cells from nor-mal cells is critical for proper analysis of disease state progression. Due to the heterogeneity of many tu-mors, however, isolation of homogenous tumor cell populations can be difficult. The CytoSelect™ Clonogenic Tumor Cell Isolation Kit uses a proprietary semisolid agar medium to facilitate colony formation by cells from solid tumors. Colonies are grown in either a 6-well plate or a 35 mm dish. The colonies are then isolated away from single cells by size filtration.

Efficient: Easily eliminates single cells from clonogenic tumor cell population

Versatile: In addition to solid tumors, has potential use in isolating tumor stem cells

Clonogenic Tumor Cell Isolation Procedure.



CytoSelect™ Leukocyte Endothelium Adhesion Assays


Fluorometric 96 Assays CBA-210


CytoSelect™ Tumor-Endothelium Adhesion Assay Fluorometric 96 Assays CBA-215

CytoSelect™ Leukocyte-Endothelium Adhesion Assay

CytoSelect™ Leukocyte-Epithelium Adhesion Assay

CytoSelect™ Leukocyte-endothelium Adhesion Assay Principle.

The CytoSelect™ Leukocyte Endothelium Adhesion Assays provide a robust system for the quantitative determination of interactions between leukocytes and endothelium. Adherent cells can be quantified on a fluorescence plate reader.

10

CELL-BASED ASSAYS Cell Adhesion

Cell Adhesion Assays

Cell adhesion is a complex mechanism involved in a variety of processes including cell mi-gration/invasion, embryogenesis, wound healing and tissue remodeling. Cells can adhere to the ECM, forming complexes with cytoskeletal components, or to the endothelium.

Our CytoSelect™ Cell Adhesion Assays quantify adhesion of cells using a microplate reader or fluorometer; no manual cell counting is required.

Recent Product Citations 1. Liu, G. et al. (2012). ICAM-1-activated Src and eNOS signal-

ing increase endothelial cell surface PECAM-1 adhesivity and neutrophil transmigration. Blood 120:1942-1952. (CBA-210)

2. Fang, Y. et al. (2012). Site-specific microRNA-92a regulation of Kruppel-like factors 4 and 2 in atherosusceptible endothe-lium. Arterioscler. Thromb. Vasc. Biol. 32:979-987. (CBA-210)

3. Curatola, A.M. et al. (2012). Dehydroepiandrosterone (DHEA) inhibition of monocyte binding by vascular endothelium is associated with sialylation of neural cell adhesion of neural cell adhesion molecule. Repr. Sciences 19:86-91. (CBA-210)

4. Wang, Y.L. et al. (2011). Innate immune function of the adher-ens junction protein p120-catenin in endothelial response to endotoxin. J. Immunol. 186:3180-3187. (CBA-210)

5. Ziemann, A. et al. (2013). CRN2 enhances the invasiveness of glioblastoma cells. Neuro Oncology 10.1093/neuonc/nos388. (CBA-215)

6. Hernandez, L. et al. (2010). Activation of NF-kB signaling by inhibitor of NF-kB Kinase ß increases aggressiveness of ovar-ian cancer. Cancer Res. 70:4005-4014. (CBA-215)

CytoSelect™ Microfluidic Biochips


CytoSelect™ 8-Channel ECM Microfluidic Biochips Microscopy 2 Chips CBA-003

10 Chips CBA-003-5

CytoSelect™ 8-Channel Endothelial Microfluidic Biochips Microscopy 2 Chips CBA-004

10 Chips CBA-004-5

CytoSelect™ 8-Channel Microfluidic Biochips provide an environment that closely mimics in vivo shear stresses, resulting in more physiologically relevant cell adhesion data. The Biochips are self-contained units containing 8 channels with inlet/outlet ports at both ends of each channel. After adding cell samples, a syringe pump set to the proper flow rate applies the appropriate shear stress to the channel.


CytoSelect™ 48-well Cell Adhesion Assay. Serum starved cells from three different cell lines were allowed to attach to the ECM-coated plate for 1 hr at 100,000 cells/well. Adherent cells were stained according to the assay protocol.

CytoSelect™ ECM Cell Adhesion Assays


CytoSelect™ 48-Well Cell Adhesion Assay, ECM Array (Contains one row each of Collagen I, Collagen IV, Fibrinogen, Fibronectin, and Laminin)

Colorimetric 48 Assays CBA-070


48 Assays CBA-071


CytoSelect™ 48-Well Cell Adhesion Assay, Collagen I Colorimetric 48 Assays CBA-052


CytoSelect™ 48-Well Cell Adhesion Assay, Collagen IV Colorimetric 48 Assays CBA-060


CytoSelect™ 48-Well Cell Adhesion Assay, Fibrinogen Colorimetric 48 Assays CBA-058


CytoSelect™ 48-Well Cell Adhesion Assay, Fibronectin Colorimetric 48 Assays CBA-050


CytoSelect™ 48-Well Cell Adhesion Assay, Laminin Colorimetric 48 Assays CBA-056


Fluorometric

The CytoSelect™ ECM Cell Adhesion Assays provide a quantitative method to measure cell adhesion. The 48-well plate is precoated with your choice of substrate. Adherent cells attach, while non-adherent cells are washed away. Adherent cells can be quantified on a standard plate reader or fluorometer.

Quantitative: Measure results in a colorimetric or fluorescence plate reader

Flexible: Uniform substrate layer of your choice of Collagen I, Collagen IV, Fibrinogen, Fibronectin, or Laminin; or choose the ECM array which contains all 5 ECM proteins

11

CELL-BASED ASSAYS Cell Adhesion

BSA

Vitronectin

Laminin

Fibronectin

Collagen IV

Collagen I

MDA-231 HT-1080 HEK293

Recent Product Citations 1. Cervera, A.M. et al. (2008). Cells silenced for SDHB expres-

sion display characteristic features of the tumor phenotype. Cancer Res. 68:4058-4067. (CBA-050 and CBA-070)

2. Lee, J. et al. (2013). Selective inhibition of prostaglandin E2 receptors EP2 and EP4 inhibits adhesion of human endometri-otic epithelial and stromal cells through suppression of integrin-mediated mechanisms. Biol. Reprod. 88:77. (CBA-057)

3. Miao, H. et al. (2008). Gene expression and functional studies of the optic nerve head astrocyte transcriptome from normal African Americans and Caucasian Americans donors. PLoS One 3(8):E2847. (CBA-060)

4. Tanoury, Z. et al. (2014). Genes involved in cell adhesion and signaling: a new repertoire of retinoic acid receptor target genes in mouse embryonic fibroblasts. J. Cell Sci. 127:521-533. (CBA-070)

5. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103. (CBA-070)

6. Mei, J. et al. (2012). Inhibition of IDO1 suppresses cyclooxy-genase-2 and matrix metalloporeinase-9 expression and de-creases proliferation, adhesion and invasion of endometrial stromal cells. Mol. Human Reprod. 10.1093/molehr/gas021. (CBA-070)

7. Kandalam, V. et al. (2011). Lack of tissue inhibitor of metallo-proteinases 2 leads to exacerbated left ventricular dysfunction and adverse extracellular matrix remodeling in response to biomechanical stress. Circulation 124:2094-2105. (CBA-070)

8. Tsukamoto, H. et al. (2010). Evaluation of anticancer activities of benzo[c]phenanthridine alkaloid sanguinarine in oral squamous cell carcinoma cell line. Anticancer Res. 31:2841-2846. (CBA-070)

9. Kim, S.W. et al. (2013). Cardiac stem cells with electrical stimu-lation improve ischaemic heart function through regulation of connective tissue growth factor and miR-378. Cardiovasc. Res. 10.1093/cvr/cvt192. (CBA-071)


CELL-BASED ASSAYS Cell Migration / Invasion


Cell Migration & Invasion Assays

Cell migration and invasion are highly integrated, multi-step processes and play important roles in the progression of various diseases including cancer, atherosclerosis and arthritis.

Our cell migration assays are provided in two formats: 2-Dimensional Gap Closure and Boyden Chamber. Each format has its own advantages and applications. Use the informa-tion below to help choose the best format for your cell migration experimental goals. Cell invasion assays are provided in the Boyden Chamber format.

Cell Migration Format Selection Guide

2D Gap Closure Assays

(p. 13) Boyden Chamber Assays

(p. 14-19)

Type of Analysis Qualitative or Quantitative Quantitative

Detection Time Endpoint or Real Time Endpoint

Detection Method Microscopy Plate Reader

Chemoattractant Gradient No Yes

Relative Sensitivity Good Fair

Adaptability to Automation Good Poor

Cell Compatibility Universal Choose pore size based on cell size

Example of Boyden Chamber Assay Principle.

Assay Principle for the Radius™ 2D Gap Closure Assays.


13 www.cellbiolabs.com [email protected]

Radius™ Cell Migration Assays (2D Gap Closure)

Example Results using 2D Gap Closure Assay.


Radius™ 24-Well Cell Migration Assay Microscopy 24 Assays CBA-125


Radius™ 24-Well Cell Migration Assay (Collagen I Coated) Microscopy 24 Assays CBA-125-COL

Radius™ 24-Well Cell Migration Assay (Fibronectin Coated) Microscopy 24 Assays CBA-125-FN

Radius™ 24-Well Cell Migration Assay (Laminin Coated) Microscopy 24 Assays CBA-125-LN

Radius™ 24-Well Cell Migration Assay (ECM Array Coated) Microscopy 24 Assays CBA-125-ECM

Radius™ 96-Well Cell Migration Assay Microscopy 96 Assays CBA-126


Radius™ 384-Well Cell Migration Assay 384 Assays CBA-127

5 x 384 Assays CBA-127-5 Microscopy

Radius™ Cell Migration Assays provide a unique al-ternative to the traditional Boyden Chamber migration assay. Radius assays allow you to measure cell mi-gration at endpoint or in real time, and are ideal for time course migration studies. Radius™ Cell Migration Assays use a cell culture plate containing a proprietary, carefully-defined bio-compatible hydrogel (Radius™ gel) spot centralized at the bottom of each well. Cells seeded in the well will attach everywhere except on the Radius gel spot, creating a cell-free zone. Once cells attach, the Ra-dius gel is removed and migration of cells across the cell-free zone begins. The gel removal step allows synchronization of a zero time point to facilitate well-to-well comparisons. With Radius™ Cell Migration Assays, there are no cell culture inserts; so you don’t need to worry about which pore size to choose for your cell type. Any ad-herent cell may be used in the assay. Radius assays are supplied in 24-well, 96-well and 384-well formats. In addition, the 24-well assays are provided with your choice of coatings for proper cell attachment: Uncoated Collagen I-coated Fibronectin-coated Laminin-coated ECM Array with 6 wells of each of the above

(uncoated, Collagen I, Fibronectin, Laminin); ideal if you are unsure which ECM protein may provide the best cell attachment

Recent Product Citations 1. Sun, J. et al. (2013). Targeting the metastasis suppressor,

NDRG1, using novel iron chelators: regulation of stress fiber-mediated tumor cell migration via modulation of the ROCK1/pMLC2 signaling pathway. Mol. Pharmacol. 83:454-469. (CBA-125)

2. Apostolos, K. et al. (2013). Increased susceptibility of melanin-concentrating hormone-deficient mice to infection with Salmonella enterica serovar Typhimurium. Infect. Immun. 81:166-172. (CBA-125)

3. Smith, K. et al. (2012). Human family with sequence similarity 60 member A (FAM60A) protein: a new subunit of the Sin3 deacety-lase complex. Mol. Cell Proteomics 11:1815-1828. (CBA-125)

4. Young, S. et al. (2012). Rapid protein kinase D1 signaling pro-motes migration of intestinal epithelial cells. Am. J. Physiol. Gas-trointest. Liver Physiol. 303:G356-G366. (CBA-125)

5. Larive, R.M. et al. (2012). The Ras-like protein R-Ras2/TC21 is important for proper mammary gland development. Mol. Biol. Cell 23:2373-2387. (CBA-125)

6. Wong, B. et al. (2013). Adrenomedullin enhances invasion of hu-man extravillous cytotrophoblast-derived cell lines by regulation of urokinase plasminogen activator expression and S-nitrosylation. Biol. Reprod. 88:34. (CBA-126)

7. Ichikawa, A. et al. (2013). CXCL10-CXCR3 enhances the develop-ment of neutrophil-mediated fulminant lung injury of viral and non-viral origin. Am. J. Respir. Crit. Care Med. 187:65-77. (CBA-126)

8. Coulouarn, C. et al. (2012). Hepatocyte-stellate cell cross-talk in the liver engenders a permissive inflammatory microenvironment that drives progression in hepatocellular carcinoma. Cancer Res. 72:2533-2542. (CBA-126)

9. Alcolea, S. et al. (2012). Interaction between head and neck squamous cell carcinoma cells and fibroblasts in the biosynthesis of PGE2. J. Lipid Res. 53:630-642. (CBA-126)


14

Boyden Chamber Assay Selection Guide

Assay Definition Cell

Types Pore Size

Insert Coating

Assay Formats

Chemotaxis (p. 15)

Migration of cells toward a chemoattractant

(chemical signal) in the cell’s surrounding environment

Neutrophils Leukocytes

3 µm None 24-Well 96-Well

Lymphocytes Monocytes

Macrophages 5 µm None

24-Well 96-Well

Fibroblasts Endothelial Cells Epithelial Cells

Tumor Cells

8 µm None 24-Well 96-Well

Astrocytes Slow-moving Cells

12 µm None 24-Well

Haptotaxis (p. 16)

Migration of cells along a gradient of cellular adhesion sites or extracellular matrix-

bound chemoattractants


8 µm

Collagen I(bottom)

24-Well

Fibronectin(bottom)

24-Well

Transmigration (p. 17)

Migration of cells through the vascular endothelium toward a

chemoattractant

Leukocytes 3 µm None 24-Well

Tumor Cells 8 µm None 24-Well

Invasion (p. 18-19)

Movement of cells through the 3D extracellular matrix into

neighboring tissues; includes ECM degradation and

proteolysis


Tumor Cells

8 µm

ECM Matrix (top)

24-Well 96-Well

Collagen I (top)

24-Well 96-Well

Laminin I (top)

24-Well 96-Well


CytoSelect™ Cell Migration and Invasion Assays (Boyden Chamber)

The Boyden Chamber has been extensively used and widely published as a tool for measuring cell migra-tion and cell invasion in vitro. Our CytoSelect™ Cell Migration and Invasion Assays use this well-cited method to quantify cell migration and invasion with no manual cell counting required. Migratory or invasive cells are quantified using a colorimetric or fluorometric plate reader. Cell migration may take on various forms and behav-iors depending on the type and location of cells. Such subclasses of cell migration include chemotaxis, hap-totaxis, and transmigration. Use the chart below to compare the various subclasses of cell migration as well as cell invasion, which will help you choose the assay best suited to your experimental goals.

Typical Well Setup for Boyden Chamber Assay.

15

CytoSelect™ Cell Migration Assays—Chemotaxis

Product Name Pore Size Detection Size Catalog Number

CytoSelect™ 24-Well Cell Migration Assay

3 µm Fluorometric 12 Assays CBA-103




8 µm





12 µm Colorimetric 12 Assays CBA-107


CytoSelect™ 96-Well Cell Migration Assay





8 µm 96 Assays CBA-106

5 x 96 Assays CBA-106-5 Fluorometric

Migration of Human Fibrosarcoma HT-1080 Cells. Cells were seeded at 30,000 cells per well of a 24-well plate and allowed to migrate toward 10% FBS for 4 hours. Migratory cells were stained (above) and quantified in a fluorescence plate reader (data not shown).

Fast Results: Visualize chemotaxis in less than 6 hours with most cell types

Flexible: Bottoms of membrane inserts are un-coated to allow use with any chemoattractant

Higher Throughput: 96-well format available for fluorescence plate readers

CytoSelect™ Cell Migration Assays are ideal for measuring chemotaxis. The kits utilize polycarbonate membrane inserts in 24-well or 96-well plates. Inserts are available with 4 different pore sizes to accommo-date a variety of cell types.

10% FBS 0% FBS


Recent Product Citations 1. Al-Wadei, M.H. et al. (2013). Gamma-amino butyric acid (GABA)

prevents the induction of nicotinic receptor-reuglated signaling by chronic ethanol in pancreatic cancer cells and normal duct epithe-lia. Cancer Prev. Res. 6:139-148. (CBA-100)

2. Chen, Z. et al. (2012). The iron chelators Cp44mT and DFO in-hibit TGF-ß-induced epithelial-mesenchymal transition via up-regulation of N-Myc downstream-regulated gene 1 (NDRG1). J. Biol. Chem. 287:17016-17028. (CBA-100)

3. Izhak, L. et al. (2010). Predominant expression of CCL2 at the tumor site of prostate cancer patients directs a selective loss of immunological tolerance to CCL2 that could be amplified in a beneficial manner. J. Immunol. 184:1092-1101. (CBA-101)

4. Kiss, J. et al. (2012). Loss of the oxygen sensor PHD3 enhances the innate immune response to abdominal sepsis. J. Immunol. 189:1955-1965. (CBA-102)

5. Verma, S. et al. (2011). Selenoprotein K knockout mice exhibit deficient calcium flux in immune cells and impaired immune re-sponses. J. Immunol. 186:2127-2137. (CBA-103)

6. Li, X. et al. (2011). Kaposi’s sarcoma-associated Herpesvirus-encoded latency-associated nuclear antigen recduces interleukin-8 expression in endothelial cells and impairs neutrophil chemo-taxis by degrading nuclear p65. J. Virol. 85:8606-8615. (CBA-104)

7. Jun, H.S. et al. (2012). Glucose-6-phosphatase-ß, implicated in a congenital neutropenia syndrome, is essential for macrophage energy homeostasis and functionality. Blood 119:4047-4055. (CBA-105)

8. Rosenblum, S. et al. (2012). Timing of intra-arterial neural stem cell transplantation after hypoxia-ischemia influences cell engraft-ment, survival, and differentiation. Stroke 43:1624-1631. (CBA-106)

9. Aftab, B.T. et al. (2011). Itraconazole inhibits angiogenesis and tumor growth in non-small cell lung cancer. Cancer Res. 71:6764-6772. (CBA-106)



CytoSelect™ Cell Migration Assays—Haptotaxis


CytoSelect™ 24-Well Cell Haptotaxis Assay, Collagen I-coated Colorimetric 12 Assays CBA-100-COL

Fluorometric 12 Assays CBA-101-COL

Colorimetric 12 Assays CBA-100-FN

Fluorometric 12 Assays CBA-101-FN CytoSelect™ 24-Well Cell Haptotaxis Assay, Fibronectin-coated

CytoSelect™ 24-well Cell Haptotaxis Assay. MDA-231 cells were seeded at 150,000 cells/well and allowed to migrate toward FBS for 4 hrs. Migratory cells, found on the bottom of the migration membrane, were stained according to the assay protocol.

Haptotaxis describes the migration of cells toward a gradient of immobilized extracellular matrix. The CytoSelect™ Cell Haptotaxis Assays are ideal for determining the migratory properties of cells. The kits utilize polycarbonate membrane inserts with an 8 µm pore size in a 24-well plate. The undersides of the inserts are coated with either Collagen or Fibronectin. The 8 µm pore size in the membrane inserts is ideal for epithelial cells, endo-thelial cells, fibroblasts, and other cells of similar size. The membrane serves as a barrier that allows discrimination of migratory cells from non-migratory cells.

Fast Results: Visualize cell haptotaxis in less than 6 hours with most cell types

Convenient: Membrane inserts pre-coated on the underside with either Collagen I or Fibronectin

Versatile: Useful with a variety of cell types including epithelial cells, endothelial cells, and fibroblasts*

*For leukocyte migration a 3 µm pore size is recommended. See our CytoSelect™ Chemotaxis Assays (previous page) or the CytoSelect™ Leukocyte Transmigration Assay (next page).

16

Recent Product Citations 1. Herrera, I. et al. (2013). Matrix metalloproteinase (MMP)-1 in-

duces lung alveolar epithelial cell migration and proliferation, protects from apoptosis, and represses mitochondrial oxygen consumption. J. Biol. Chem. 288:25964-25975. (CBA-100-COL)

2. Niccoli, S. et al. (2012). The Asian-American E6 variant protein of human papillomavirus 16 alone is sufficient to promote immor-talization, transformation, and migration of primary human fore-skin keratinocytes. J. Virol. 86:12384-12396. (CBA-110-COL)

3. Kamiya, K. et al. (2007). Protein Kinase C delta activated adhe-sion regulates vascular smooth muscle cell migration. J. Surg. Res. 141:91-96. (CBA-100-COL)

Assay Principle for the CytoSelect™ Cell Haptotaxis Assay.

BSA Collagen I Fibronectin

0% FBS

0.5% FBS



17

CytoSelect™ Cell Migration Assays—Transmigration


CytoSelect™ Leukocyte Transmigration Assay Fluorometric 24 Assays CBA-212

Fluorometric 24 Assays CBA-216 CytoSelect™ Tumor Transendothelial Migration Assay

Pore Size

3 µm

8 µm

Cancer cell transmigration, particularly extravasation, is an important step in cancer metastasis. The CytoSe-lect™ Cell Transmigration Assays provide a robust system for the quantitation of transmigrations and interac-tions between endothelium and cancer cells. Migratory cells are quantified via fluorometer.

Recent Product Citations 1. Fava, G. et al. (2008). Leptin enhances cholangiocarcinoma cell

growth. Cancer Res. 68:6752-6761. (CBA-212) 2. Park, G.B. et al. (2014). The Epstein-Barr Virus causes epithelial

-mesenchymal transition in human corneal epithelial cells via Syk/Src and Akt/ERK signaling pathways. Invest. Ophthalmol. Vis. Sci. 55:1770-1779. (CBA-216)

3. Xu, Z. et al. (2010). Role of pancreatic stellate cells in pancreatic cancer metastasis. Am. J. Pathol. 177:2585-2596. (CBA-216)

Assay Principle for the CytoSelect™ Leukocyte Transmigration Assay.

The Leukocyte Adhesion and Transmigration Cascade.


0

40

80

120

160

200

0 10,000 20,000 30,000 40,000 50,000

Cell Number

RF

U

Quantitation of Human Monocytic THP-1. LeukoTracker™ labeled THP-1 cells were titrated in 1X PBS, then lysed with 2X lysis buffer. Fluorescence was quantified as described in the assay protocol.

CytoSelect™ Cell Invasion Assays

Quantitative: Measure results in a colorimet-ric or fluorescence plate reader

Flexible: Uniform protein matrix layer of your choice of basement membrane (from mouse tumor cells), Collagen I, or Laminin I

Versatile: Characterize both the invasive and migratory properties of your cells with a Cell Migration / Invasion Combo Kit (next page)

CytoSelect™ Cell Invasion Assay Principle.

Human Fibrosarcoma HT-1080 Laminin I Cell Invasion. HT-1080 and NIH3T3 (negative control) were seeded at 200,000 cells/well and allowed to invade toward FBS for 24 hrs. Invasive cells on the membrane bottom were stained (top and center) and quantified at OD 560nm after extraction (data not shown).

Tumor cell invasion into surrounding normal tissue contributes to the morbidity of cancers. The CytoSe-lect™ Cell Invasion Assays use precoated inserts to assay invasive properties of tumor cells in 24-well or 96-well plates. The coated layer serves to distinguish invasive cells from non-invasive cells. Plates are pre-coated with either basement membrane matrix (from EHS mouse sarcoma cells), Collagen I or Laminin I.

18


Recent Product Citations 1. Gradilone, S. et al. (2013). HDAC6 inhibition restores ciliary

expression and decreases tumor growth. Cancer Res. 73:2259-2270. (CBA-110)

2. Nam, H. et al. (2013). Antitumor activity of saracatinib (AZD0530), a c-Src/Abl kincase inhibitor, alone or in combina-tion with chemotherapeutic agents in gastric cancer. Mol. Can-cer Ther. 12:16-26. (CBA-110)

3. DiNatale, B. et al. (2012). Ah receptor antagonism represses head and neck tumor cell aggressive phenotype. Mol. Cancer Res. 10:1369-1379. (CBA-110)

4. Citterio, C. et al (2012). The Rho exchange factors Vav2 and Vav3 control a lung metastasis-specific transcriptional program in breast cancer cells. Sci. Signal. 5:ra71. (CBA-110)

5. Nakayama, K. et al. (2013). cAMP-response element-binding protein (CREB) and NFkB transcription factors are activated during prolonged hypoxia and cooperatively regulate the induc-tion of matrix metalloproteinase MMP1. J. Biol. Chem. 288:2584-22595. (CBA-112)

6. Desai, S.D. et al. (2012). ISG15 disrupts cytoskeletal architec-ture and promotes motility in human breast cancer cells. Exp. Biol. Med. 237:38-49. (CBA-112)

7. Ziemann, A. et al. (2013). CRN2 enhances the invasiveness of gliobastoma cells. Neuro Oncology 10.1093/neuonc/nos388. (CBA-112-COL)

8. Liu, X. et al. (2013). Antiproliferative, antiinvasive, and proapoptotic activity of folate receptor alpha-targeted liposomal doxorubicin in nonfunctional pituitary adenoma cells. Endocri-nology 154:1414-1423. (CBA-112-COL)


CytoSelect™ Cell Invasion Assays, continued


CytoSelect™ 24-Well Cell Invasion Assay, Basement Membrane Colorimetric 12 Assays CBA-110


CytoSelect™ 24-Well Cell Invasion Assay, Collagen I Colorimetric 12 Assays CBA-110-COL

Fluorometric 12 Assays CBA-111-COL

Colorimetric 12 Assays CBA-110-LN

Fluorometric 12 Assays CBA-111-LN

CytoSelect™ 96-Well Cell Invasion Assay, Basement Membrane Fluorometric 96 Assays CBA-112

CytoSelect™ 96-Well Cell Invasion Assay, Collagen I Fluorometric 96 Assays CBA-112-COL

CytoSelect™ 96-Well Cell Invasion Assay, Laminin I Fluorometric 96 Assays CBA-112-LN

CytoSelect™ 24-Well Cell Invasion Assay, Laminin I

CytoSelect™ Cell Migration / Invasion Assay Combo Kits

Effects of Cytochalasin D on Invading Cells using the CytoSelect™ 24-well Cell Invasion Assay (CBA-110). HT-1080 and NIH3T3 cells (negative control) were seeded at 300,000 cells/well and allowed to invade toward 10% FBS for 24 hrs, in the presence or absence of 2 µM Cytochalasin D. Invasive cells, on the bottom of the invasion membrane, were stained (left) and then quantified at OD 560 nm after ex-traction using a standard plate reader (right).

Our CytoSelect™ Cell Migration / Invasion Assay Combo Kits allow you to characterize both the migratory and invasive properties of your cells. Each 24-well combo kit provides sufficient reagents to perform 12 migration plus 12 invasion assays, while the 96-well combo kit allows you to perform 96 migration plus 96 invasion assays. The invasion plate provided contains basement membrane-coated inserts.

19

NIH3T3 HT-1080

0

0.4

0.8

1.2

1.6

2

NIH3T3 HT-1080 HT-1080 +Cytochalasin D

OD

560

nm


Recent Product Citations 1. Majid, S. et al. (2013). miRNA-34b inhibits prostate cancer through demethylation, active chromatin modification, and AKR pathways. Clin.

Cancer Res. 19:73-84. (CBA-100-C) 2. Shin, S.Y. et al. (2012). Transcriptional regulation of the interleukin-11 gene by oncogenic Ras. Carcinogenesis 10.1093/carcin/bgs297.

(CBA-100-C) 3. Gobeil, S. et al. (2008). A genome-wide shRNA screen identifies GAS1 as a novel melanoma metastasis suppressor gene. Genes Dev.

22(21):2932-2940. (CBA-101-C) 4. Axlund, S.D. et al. (2010). HOXC8 inhibits androgen receptor signaling in human prostate cancer cells by inhibiting SRC-3 recruitment to

direct androgen target genes. Mol. Cancer Res. 8:1643-1655. (CBA-106-C)

Product Name Pore Size Detection Size Catalog Number

CytoSelect™ 24-Well Cell Migration / Invasion Combo Kit Colorimetric 2 x 12 Assays CBA-100-C

Fluorometric 2 x 12 Assays CBA-101-C

CytoSelect™ 96-Well Cell Migration / Invasion Combo Kit 8 µm Fluorometric 2 x 96 Assays CBA-106-C

8 µm


20


CytoSelect™ 24-Well Wound Healing / Cell Migration Assay

Highly Accurate: More consistent results well-to-well compared to homemade scratch assays

Versatile: Measure cell migration, cell prolifera-tion, and wound closure

Inert Material: No residues from inserts to impede cell migration or proliferation

Compared to traditional scratch assays, our CytoSe-lect™ 24-Well Wound Healing Assay provides a more consistent method to measure cell migration across a “wound field” gap in vitro. Proprietary treated inserts generate a consistently defined 0.9mm gap between the cells. Cells can then be treated and monitored for proliferation or migration across the wound field by imaging samples at fixed time points or time-lapse microscopy.


CytoSelect™ 24-Well Wound Healing Assay 24 Assays CBA-120

5 x 24 Assays CBA-120-5 Microscopy

Wound Closure of STO Cells. STO cells (mouse MEF) were cul-tured in the provided plate with inserts in place for 24 hours until a monolayer formed. Inserts were then removed to begin the assay. Cells were monitored at various time points and stained according to the assay protocol. CytoSelect™ 24-well Wound Healing Assay Principle.

0%

50%

100%

Percent Wound Closure


Recent Product Citations 1. Cui, X.G. et al. (2013). Response gene to complement 32 defi-

ciency causes impaired placental angiogenesis in mice. Cardio-vasc. Res. 10.1093/cvr/cvt121.

2. Gutschner, T. et al. (2013). The noncoding RNA MALAT1 is a critical regulator of the metastasis phenotype of lung cancer cells. Cancer Res. 73:1180-1189.

3. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103.

4. Mao, R. et al. (2012). Circadian gating of epithelial-to-mesenchymal transition in breast cancer cells via melatonin-regulation of GSK3ß. Mol Endrocrinol. 26:1808-1820.

5. Hirata, H. et al. (2012). MicroRNA-1826 targets VEGFC, beta-catenin (CTNNB1) and MEK1 (MAP2K1) in human bladder can-cer. Carcinogenesis 33:41-48.

21

CELL-BASED ASSAYS Cell Co-Culture



CytoSelect™ 24-Well Cell Co-Culture System Microscopy 24 Assays CBA-160

CytoSelect™ 24-Well Cell Co-Culture System

Traditional methods of co-culture usually involve one of the following methods: 1. One cell type is cultured to form a monolayer,

followed by seeding of a second cell type directly over the monolayer. This is a common method when feeder cells are used to maintain stem cells in an undifferentiated state. However, it is not useful when studying the effects of one cell on the other because the first cell is obscured from view by the second.

2. A Boyden Chamber (see page 12 for details) is used to culture one cell type above the mem-brane and a second cell type below the mem-brane. This system allows a separation between the two cells, but does not allow for direct cell-to-cell contact which may reduce its efficacy for cer-tain applications.

The CytoSelect™ Cell Co-Culture System provides a unique platform for direct contact between two cell types in one well. A proprietary molded plastic insert creates a cell-free zone in the center of a 24-well cell culture-treated plate. The first cell type is seeded in the area around the insert. Once the cells form a monolayer, the insert is removed and the second cell is seeded.

The culture of two cell lines together is advantageous for studying a variety of applications: Cell-cell interactions Cell activation Cellular differentiation Maintaining stem cell pluripotency Various effects of secreted factors from one cell type on a second cell type

Protocol for the CytoSelect™ 24-Well Co-Culture System. Cell type #1 is seeded with the Co-Culture insert in place. After cells form a monolayer, the insert is removed and cell type #2 is seeded.

CELL-BASED ASSAYS Cell Health

22

CytoSelect™ Cell Viability and Cytotoxicity Assay (Live/Dead)


CytoSelect™ Cell Viability and Cytotoxicity Assay Kit Colorimetric / Fluorometric

96 Assays CBA-240

Cell viability characteristics include cellular metabolic activity and cell membrane integrity. Our CytoSe-lect™ Cell Viability and Cytotoxicity Assay provides both a colorimetric and fluorometric format for moni-toring cell viability via metabolic activity. Live cells are detected with MTT (colorimetric detection) or Calcein AM (fluorometric); dead cells are detected with EthD-1 reagent (fluorometric). All 3 detection reagents are included, as well as Saponin, a cell death initiator. Cells may be treated with compounds or agents that affect cell viability. This kit is suitable for eukaryotic cells, not bacteria or yeast.

Viability of Human Foreskin Fibroblasts. BJ-TERT cells were seeded at 50,000 cells/well and allowed to culture for 24 hours. Cells were then treated with and without Saponin. All cells were then stained with Calcein AM and EthD-1. Top: Cells without Saponin treatment. Bottom: Cells with Saponin treatment. Left: Calcein AM staining. Right: EthD-1 staining.

Versatile: Detect by microscopy, colorimetric or fluorescence plate reader, or flow cytometry

Quantitative: Measure live and dead cells on a fluorescence plate reader; live cells may also be quantified on a standard microplate reader


Recent Product Citation Kim, E.Y. et al. (2012). Sustained activation of N-methyl-D-aspartate receptors in podocytes leads to oxidative stress, mobili-zation of transient receptor potential canonical 6 channels, nuclear factor of activated T cells activation, and apoptotic cell death. Mol. Pharmacol. 82:728-737.

CytoSelect™ LDH Cytotoxicity Assay


CytoSelect™ LDH Cytotoxicity Assay Kit Colorimetric 960 Assays CBA-241

Loss of cell membrane integrity is one of the hall-marks of cytotoxicity. Upon cell death, lactate dehy-drogenase (LDH) is released from the cytoplasm through the damaged membrane. Our CytoSelect™ LDH Cytotoxicity Assay provides a convenient plate-based method for testing cytotoxicity based on LDH release. In this assay, cells are cul-tured in a 96-well plate with and without the com-pound to be tested. LDH released into the media from cells converts a lactate substrate to pyruvate and generates NADH. In the presence of NADH, the col-orimetric dye WST-1 is converted to a formazan that generates an orange color. Detection is performed in a standard colorimetric plate reader.

LDH Release from HEK 293 Cells. 20,000 cells/well were cultured for 24 hours. After adding various concentrations of Triton X-100, the LDH Cytotoxicity Assay Reagent was added followed by a 30 minute incubation at 37ºC and 5% CO2.

CytoSelect™ Anoikis Assays


CytoSelect™ 24-Well Anoikis Assay Colorimetric / Fluorometric

24 Assays CBA-080

CytoSelect™ 96-Well Anoikis Assay Colorimetric / Fluorometric

96 Assays CBA-081

Anoikis of Human Fibroblast BJ-TERT Cells. 50,000 cells/well were seeded in a control plate (left) and a Poly-HEMA coated plate (right) and cultured for 24 hours. Cells on the control plate were stained with Calcein AM. Cells on the Poly-HEMA coated plate were stained with EthD-1.

Anoikis is defined as death of adherent cells due to loss of adhesion to the extracellular matrix. Our Anoikis Assays allow you to quantify and monitor an-chorage-dependent cell death using a precoated plate. Live cells can be viewed under a microscope and quantified on a plate reader by MTT (colorimetric) or Calcein AM (fluorometric), both in-cluded with the kit. Dead cells are detected with an EthD-1 reagent.

Versatile: Detect live and dead cells by micros-copy, fluorescence, or flow cytometry

Quantitative: Measure live and dead cells on a fluorescence plate reader; live cells may also be quantified on a standard microplate reader

Cellular Senescence Assays


Cellular Senescence Assay Kit (SA -gal Staining) Light Microscopy 50 Assays CBA-230


96-Well Cellular Senescence Assay (SA -gal Activity) Fluorometric Plate Reader

120 Assays CBA-231


Flow Cytometry / Fluorescence Microscopy

10 Assays CBA-232

5 x 10 Assays CBA-232-5 Quantitative Cellular Senescence Assay (SA -gal)

Senescence Associated (SA) -galactosidase is a common biochemical marker of cellular senescence. Cells expressing such markers have been identified in vivo in tissues. SA ß-Gal catalyzes hydrolysis of X-gal, which produces a blue color in senescent cells. We offer three kit formats to test cellular senescence via SA-ß-galalactosidase activity: Our ß-Gal Staining Kit allows you to visualize senescence by standard light microscope. Our Quantitative Cellular Senescence Assay measures senescence in cells cultured in a 35mm dish by

either flow cytometry or fluorescence microscopy Our 96-Well Cellular Senescence Assay provides a higher throughput assay in a fluorescence plate reader.

Recent Product Citations 1. Lee, H.W. et al. (2013). Tpl2 kinase impacts tumor growth

and metastasis of clear cell renal cell carcinoma. Mol. Can-cer Res. 11:1375-1386. (CBA-080)

2. Sisto, M. et al. (2009). Fibulin-6 expression and anoikis in human salivary gland epithelial cells: implications in Sjogren's syndrome. Int. Immunol. 21:303-311. (CBA-080)

3. Liu, H. et al (2008). Cysteine-rich protein 61 and connective tissue growth factor induce de-adhesion and anoikis of reti-nal pericytes. Endocrinology 149:1666-1677. (CBA-080)



Recent Product Citation Malhotra, D. et al. (2010). Global mapping of binding sites for Nrf2 identifies novel targets in cell survival response through ChIP-Seq profil-ing and network analysis. Nucleic Acid Res. 10.1093/nar/gkq212. (CBA-231)



CytoSelect™ MTT Cell Proliferation Assay


CytoSelect™ MTT Cell Proliferation Assay Colorimetric 960 Assays CBA-252

Cell proliferation is easily measured by the addition of a variety of dyes that produce a visible color that can be correlated with the number of cells. Our CytoSelect™ MTT Cell Proliferation Assay provides a simple method to measure proliferation of cells. The cell-permeable MTT dye is added directly to cultured cells followed by a de-tergent solution. Quantitation is performed using a standard microplate reader at 540-570 nm.

CytoSelect™ WST-1 Cell Proliferation Assay Reagent

Our CytoSelect™ WST-1 Cell Proliferation Assay provides a similar method to our MTT Cell Proliferation Assay, but with a single reagent format that does not require a detergent solubilization step. Quantitation is performed using a standard microplate reader at 450 nm.


CytoSelect™ Cell Proliferation Assay Reagent, Fluorometric Fluorometric 960 Assays CBA-250

CytoSelect™ Fluorometric Cell Proliferation Assay Reagent


CytoSelect™ WST-1 Cell Proliferation Assay Reagent Colorimetric 960 Assays CBA-253

Human HEK 293 Cell Density. Cells were seeded at various den-sities in triplicate and allowed to culture for 24 hours. Cells were then treated with the CytoSelect™ Cell Proliferation Assay Reagent for 6 hours at 37ºC and 5% CO2.

Cell proliferation is easily measured by the addition of a variety of dyes that can be correlated with the num-ber of cells. Various dyes producing a visible color are available to measure proliferation rates, but fluorometric dyes are often more sensitive and may be a superior choice for researchers with access to a fluorescence-based microplate reader. Our CytoSelect™ Cell Proliferation Assay Reagent (Fluorometric) provides a simple, single reagent method to measure proliferation of cells. The fluoro-metric dye is added directly to cultured cells. Upon entering metabolically active live cells, the non-fluorescent dye is converted to a bright red fluores-cent form. Quantitation is performed using a fluores-cence plate reader with excitation at 560 nm and emission at 590-600 nm. This reagent is versatile and can be used with a wide variety of cell types including cultured mammalian and piscine cells, bacteria, yeast, fungi, and protozoa.



CytoSelect™ BrdU Cell Proliferation ELISA Kit


CytoSelect™ BrdU Cell Proliferation ELISA Kit Colorimetric 96 Assays CBA-251

Assay Principle for the CytoSelect™ BrdU Cell Proliferation ELISA Kit.

BrdU is a thymidine analog that can incorporate into newly synthesized DNA strands of actively proliferating cells. Our CytoSelect™ BrdU Cell Proliferation ELISA Kit provides a convenient plate-based method to measure this incorporation. Once the BrdU is incorporated into the DNA, cells are fixed and DNA is denatured. Incorpo-rated BrdU can be quantified in the denatured DNA by an anti-BrdU antibody. The absorbance readings at 450 nm can be directly correlated to cell proliferation.

CytoSelect™ Proliferating Cell Nuclear Antigen (PCNA) ELISA Kit


CytoSelect™ Proliferating Cell Nuclear Antigen (PCNA) ELISA Kit Colorimetric 96 Assays CBA-254

PCNA Detection in HeLa Whole Cell Lysates. Whole cell lysates were prepared in a RIPA lysis buffer. Protein concentrations were determined by BCA protein assay.

Proliferating Cell Nuclear Antigen (PCNA) acts as a processivity factor for DNA polymerase by associat-ing with various proteins involved in DNA replication. It is also associated with chromatin remodeling and cell cycle control, and it is often used as a marker of cell proliferation. Our CytoSelect™ PCNA ELISA Kit provides a con-venient plate-based method to quantify PCNA levels in nuclear or whole cell extracts.

Sensitive: Detect PCNA as low as 12.5 ng/mL Versatile: Measure PCNA levels from human,

mouse, or rat whole cell lysates or nuclear extracts Quantitative: Measure results in a colorimetric

plate reader against a provided PCNA standard

CELL-BASED ASSAYS Cell Hypoxia


HIF-1 Alpha DNA Binding Activity Assay Kit

Detection Specificity of HIF-1 Alpha. HeLa cells were incubated in the presence or absence of 0.2 mM deferoxamine mesylate (DFO) for 4 hours at 37ºC. Nuclear extracts were prepared using the Nuclear/Cytosolic Fractionation Kit (#AKR-171). 100 pmol of non-biotinylated wild type or mutated HRE double stranded com-petitor oligos were added to the Complete DNA Binding Buffer just prior to inclusion in the assay.


HIF-1 Alpha DNA Binding Activity Assay Kit Colorimetric 96 Assays CBA-282

Cell hypoxia, or low oxygen condition, is a normal physiological response to certain body stressors such as high altitudes, but it can also be a symptom of pathological conditions and is often used as a marker for tumor cells. In response to hypoxic conditions, the hypoxia-inducible factor 1 transcriptional activator complex (HIF-1) plays a role in activating several hy-poxia-responsive genes such as erythropoietin and VEGF. During hypoxia, the alpha subunit of HIF-1 accumulates and translocates from the cytosol to the nucleus, where it dimerizes with the beta subunit and becomes transcriptionally active. It then binds tran-scriptional coactivators to induce gene expression. The HIF-1 Alpha DNA Binding Activity Assay Kit de-tects activated HIF-1 in an ELISA format. Active HIF-1 complex is captured on a double-stranded oligo containing a hypoxic response element (HRE) that is attached to the plate. Detection is then performed with a primary antibody followed by an HRP-conjugated secondary antibody. The assay will detect HIF-1 complexes from human, mouse or rat protein samples.


HIF-1 Alpha Sandwich ELISA Kit Colorimetric 96 Assays CBA-280

HIF-1 Alpha Cell Based ELISA Kit Chemiluminescent 96 Assays CBA-281

HIF-1 Alpha ELISA Kits

Detection of Nuclear HIF-1 Alpha with the HIF-1 Alpha Sand-wich ELISA Kit. HeLa cells were incubated in the presence or absence of 0.2 mM DFO for 4 hours at 37ºC. HIF-1 Alpha levels were measured in untreated (blue bars) and treated (red bars) nuclear extracts according to the Assay Protocol.

Our HIF-1 Alpha ELISA Kits provide a convenient method for detection and quantitation of human, mouse, or rat HIF-1 Alpha in cells or tissues. Two ELISA kit formats are available: The HIF-1 Alpha Sandwich ELISA Kit detects HIF

-1 Alpha in any protein sample including tissue homogenates, whole cell lysates, or nuclear ex-tracts. Samples are added to an anti-HIF-1 Alpha antibody coated plate. Quantitation of unknown samples is performed by comparison of the OD values to those of a known standard.

The HIF-1 Alpha Cell Based ELISA Kit allows the detection of HIF-1 Alpha levels in intact cells. Cells are seeded in a tissue culture treated plate suitable for reading in a 96-well plate-based lumi-nometer. Cells are fixed and permeabilized to allow detection with the anti-HIF-1 antibody. De-tection is performed by chemiluminescence.

CELL-BASED ASSAYS Adipogenesis


CytoSelect™ 96-well Adipogenesis Assay Kit


CytoSelect™ 96-Well Adipogenesis Assay Kit Colorimetric / Fluorometric

200 Assays CBA-290

Staining of 3T3-L1 Cells with Oil Red O. 20,000 cells/well of preadipocyte 3T3-L1 cells were seeded overnight in a 96-well plate. Cells were uninduced (top) or induced (bottom) for 7 days and stained with Oil Red O colorimetric stain according to the Assay Protocol.

Staining of 3T3-L1 Cells with Nile Red Fluorescent Stain. 20,000 cells/well of preadipocyte 3T3-L1 cells were seeded over-night in a 96-well plate. Cells were uninduced (top) or induced (bottom) for 7 days and stained with Nile Red Fluorescent Stain according to the Assay Protocol.

The ability to regulate the cell cycle and differentiation of adipocytes is important to the understanding of obesity. Adipogenesis is the process in which preadipocytes develop into mature adipocytes in a multistep process that requires the sequential activation of numerous transcription factors. The 3T3-L1 cell line is the best character-ized model for adipogenesis in vitro. 3T3-L1 cells display a fibroblast-like phenotype when grown under normal conditions. However, when treated with a combination of IBMX, insulin, and dexamethasone, these cells un-dergo terminal differentiation resulting in a more rounded phenotype and the formation of intracellular lipid drop-lets. The CytoSelect™ 96-Well Adipogenesis Assay quantitatively measures lipid droplet accumulation in cultured cells of the 3T3-L1 model. Quantitation is performed either in a standard colorimetric plate reader with Oil Red O stain, or in a fluorescence plate reader with Nile Red fluorometric stain.

CELL-BASED ASSAYS Phagocytosis


CytoSelect™ 96-Well Phagocytosis Assays

Phagocytosis may be assayed by measuring the en-gulfing of a cell “substrate” such as an erythrocyte(RBC) or Zymosan particle. Traditional phagocytosis assays involve manually counting the engulfed sub-strates under a microscope. This process is tedious and time-consuming, can be somewhat inaccurate, and is not amenable to high throughput. CytoSelect™ 96-Well Phagocytosis Assays are more accurate, high-throughput alternatives to the standard phagocytosis assay. The assays may be adapted for use in 48-well and 24-well plates if desired.


CytoSelect™ 96-Well Phagocytosis Assay (E. coli) Colorimetric 96 Assays CBA-222

CytoSelect™ 96-Well Phagocytosis Assay (Red Blood Cell) Colorimetric 96 Assays CBA-220


5 x 96 Assays CBA-224-5 CytoSelect™ 96-Well Phagocytosis Assay (Zymosan)

Assay Principle for the CytoSelect™ 96-Well Phagocytosis Assay (Zymosan).

Particle Engulfment with the CytoSelect™ 96-Well Phagocytosis Assay (Zymosan).

Recent Product Citations 1. Lee, J.K. et al. (2010). Regulator of G-protein signaling-10 nega-

tively regulates NF-kB in microglia and neuroprotects dopa-minergic neurons in hemiparkinsonian rats. J. Neurosci. 31:11879-11888. (CBA-220)

2. Winnicka, B. et al. (2010). CD13 is dispensible for normal hema-topoiesis and myeloid cell functions in the mouse. J. Leukoc. Biol. 88(2):347-359. (CBA-220)

3. Hamilton, C.M. et al. (2009). Fasciola hepatica tegumental anti-gen suppresses dendritic cell maturation and function. Infect. Immun. 77:2488-2498. (CBA-220)

4. Dowling, D.J. et al. (2009). Major secretory antigens of the helminth Fasciola hepatica activate a suppressive dendritic cell phenotype that attenuates Th17 cells but fails to activate Th2 immune responses. Infect. Immun. 78:793-801. (CBA-220)

5. Pierce, L.M. et al. (2012). Effect of heavy metal tungsten alloy particles on oxidative product formation and phagocytosis by lung macrophages. Am. J. Respir. Crit. Care Med. 185:A4666. (CBA-224)

6. Polancec, D.S. et al. (2012). Azithromycin drives in vitro GM-CSF/IL-4-induced differentiation of human blood monocytes toward dendritic-like cells with regulatory properties. J. Leukoc. Biol. 91:229.-243. (CBA-224)

Highly Accurate: Eliminates manual counting High Throughput: 96-well plate format Quantitative: Measure OD in a standard microplate

reader Flexible: Choose from 3 substrates: E. coli, Zymo-

san particles, or red blood cells*

*Red blood cells are not provided in the kit. Fresh RBCs should be obtained immediately prior to running the assay. E. coli and Zymosan particles are provided in their respective kits.

CELL-BASED ASSAYS Cell Contraction and Angiogenesis


Endothelial Tube Formation (In Vitro Angiogenesis) Assay


Endothelial Tube Formation Assay (In Vitro Angiogenesis) Light Microscopy 50 Assays CBA-200

Recent Product Citations 1. Yu, J.G. et al. (2013). Baroreflex deficiency hampers angiogene-

sis after myocardial infarction via acetylcholine-alpha7-nicotinic ACh receptor in rats. Eur. Heart J. 34:2412-2420.

2. Bid, H. et al. (2013). Dual targeting of the Type 1 insulin-like growth factor receptor and its ligands as an effective antiangio-genic strategy. Clin. Cancer Res. 19:2984-2994.

3. Cai, C. et al. (2012). SIVmac239-Nef downregulates cell surface expression of CXCR4 in tumor cells and inhibits proliferation, migration and angiogenesis. Anticancer Res. 23:2759-2768.

4. Bid, H. et al. (2012). Potent inhibition of angiogenesis by the IGF-1 receptor-targeting antibody SCH717454 is reversed by IGF-2. Mol. Cancer Ther. 11:649-659.

For angiogenesis to occur, endothelial cells must es-cape their stable location and break through the basement membrane. These cells proliferate to form new blood vessels. Our Endothelial Tube Formation Assay provides an easy, robust system to assess angiogenesis in vitro. The assay uses an ECM gel matrix derived from mouse sarcoma cells; this matrix very closely resembles an in vivo basement mem-brane environment.


Cell Contraction Assay Light Microscopy 24 Assays CBA-201

Collagen-Based Cell Contraction Assay

Assay Principle for the Collagen-Based Cell Contraction Assay.

Recent Product Citations 1. Kotio, K.U. et al. (2011). Implication of microRNAs in atrial natri-

uretic peptide and nitric oxide signaling in vascular smooth mus-cle cells. Am. J. Physiol. Gastrointest. Liver Physiol. 301: C929-C937.

2. Schell, C. et al. (2010). 15-deoxy-delta12-14-prostaglandin-J2 induces hypertrophy and loss of contractility in human testicular peritubular cells: implications for human male fertility. Endocri-nology 151:12571268.

Wound healing is comprised of epithelialization, con-nective tissue deposition, and contraction. The con-traction process is believed to be mediated by spe-cialized fibroblasts (myofibroblasts). 3D collagen gels have been widely used in fibroblast contraction stud-ies. Our Cell Contraction Assay provides a simple system to assess cell contractivity and to screen for cell con-traction mediators. The system uses a 3D collagen matrix to measure changes in the collagen gel size. An optional contraction inhibitor is provided.

HUVEC Tube Formation on ECM Gel. HUVEC cells from a stan-dard tissue culture plate were incubated on an ECM gel. After sev-eral hours tube formation can be visualized under a light micro-scope.

CELL-BASED ASSAYS Autophagy

30

GFP-LC3 Expression Vectors


pCMV-GFP-LC3 Expression Vector 100 µL CBA-401

pSMPUW-GFP-LC3 Lentiviral Expression Vector 10 µg LTV-801

10 µg RTV-801 pMXs-GFP-LC3 Retroviral Expression Vector


Recent Product Citations 1. Chen, W. et al. (2012). Andrographolide induces autophagic cell death in human liver cancer cells through cyclophilin D-mediated mito-

chondrial permeability transition pore. Carcinogenesis 10.1093/carcin/bgs264. (CBA-401) 2. Cina, D.P. et al. (2012). Inhibition of MTOR disrupts autophagic flux in podocytes. J. Am. Soc. Nephrol. 23:412-420. (CBA-401) 3. Tu, S.P. et al. (2011). IFN-gamma inhibits gastric carcinogenesis by inducing epithelial cell autophagy and T-cell apoptosis. Cancer Res.

71:4247-4259. (CBA-401)

MAP LC3 is the most published autophagosome marker protein. LC3 associates to the inner and outer limiting membranes of the auto-phagosome. There are two forms of LC3 visible by immunoblot: LC3I which is found in the solu-ble fraction, and LC3II which is found in the membrane fraction. The proportion of LC3II in-creases during autophagy. Our GFP-LC3 expression vectors are conven-ient tools for the study of autophagy. These vec-tors are available in three formats: mammalian, lentiviral, and retroviral expression vectors. Each vector contains a GFP reporter gene. In addi-tion, a GFP control plasmid is provided at no additional charge.

STEM CELL RESEARCH Induced Pluripotent Stem Cells

iPS Cell Reprogramming

Reprogramming of adult cells into induced pluripotent stem cells (iPS) has provided an im-portant new vehicle to facilitate stem cell research. Recent studies have shown that this may be accomplished by the introduction of key genes into somatic cells by transduction with various viral vectors or transfection of plasmids. Retroviral and lentiviral vectors appear to achieve among the highest levels of efficiency of iPS cell generation. We offer an extensive collection of vectors for iPS cell reprogramming.

32

Retroviral Vectors and Packaging Cells for iPS Cell Generation

Our iPS retroviral vectors are constructed from the pMXs vector backbone developed by Dr. Toshio Kitamura at the University of Tokyo.* Each vector contains one of 6 factors shown to help reprogram adult fibroblasts into iPS cells. Both human and mouse genes are available individually or in sets. Separate retroviral vectors are available for p53 shRNA, which has been shown to potentially increase the efficiency of iPS cell generation. Platinum Retroviral Packaging Cells provide an easy way to produce high-titer retroviruses from these stem cell plasmids. For additional information on these cell lines please see p. 64.

Target Name Vector Backbone Catalog Number

Oct-3/4 pMXs RTV-705

Sox2 pMXs RTV-706

c-Myc pMXs RTV-707

Klf4 pMXs RTV-708

NANOG pMXs RTV-711

Lin28 pMXs RTV-712

Set of 4 vectors (Oct-3-4, Sox2, c-Myc, Klf4)

pMXs RTV-705-C

Set of 6 vectors (Oct-3-4, Sox2, c-Myc, Klf4, NANOG, Lin28)

pMXs RTV-711-C

p53 shRNA pRetro RTV-400

Mouse iPS Vectors



Sox2 pMXs RTV-702

c-Myc pMXs RTV-703

Klf4 pMXs RTV-704

NANOG pMXs RTV-709

Lin28 pMXs RTV-710

Set of 4 vectors (Oct-3-4, Sox2, c-Myc, Klf4)

pMXs RTV-701-C

Set of 6 vectors (Oct-3-4, Sox2, c-Myc, Klf4, NANOG, Lin28)

pMXs RTV-709-C


Human iPS Vectors


Platinum-E Retroviral Packaging Cell Line, Ecotropic >3 x 106 cells RV-101

Platinum-A Retroviral Packaging Cell Line, Amphotropic >3 x 106 cells RV-102

Platinum-GP Retroviral Packaging Cell Line, Pantropic >3 x 106 cells RV-103

pCMV-VSV-G Packaging Vector (for use with Platinum-GP cells) 10 µg RV-110

Retroviral Packaging Cell Lines

*Kitamura, T. et al. (2003). Exp. Hematol. 31:1007-1014.


Induced Pluripotent Stem Cells

Polycistronic Vectors for iPS Cell Generation

STEM CELL RESEARCH

33


pLentG-KOSM Polycistronic Lentiviral Vector (Mouse genes) 100 µL LTV-700

pRetroG-OKSM Polycistronic Retroviral Vector (Human genes) 100 µL RTV-700

Our Polycistronic Viral Vectors provide a convenient way to generate iPS cells. The defined stem cells factors Klf4, Oct-3/4, Sox2 and c-Myc are in-frame fused into a single open reading frame (ORF) by self-cleaving 2A peptides. The transcription factor ORF is followed by IRES-GFP as a reporter to verify viral transduction into your target cell. Efficiencies of iPS generation are typically higher compared to transduc-tion of four separate viruses each containing a single gene. Two vectors are available: pLentG-KOSM is a lentiviral vector containing

mouse sequences pRetroG-OKSM is a retroviral vector containing

human sequences

More Efficient: Up to 10-fold higher efficiency compared to multi-virus transduction, and 500-fold compared to non-viral methods

Reporter Convenience: GFP reporter gene helps to monitor viral transduction

Open Reading Frame of pLentG-KOSM Lentiviral Vector.

Characterization of iPS Cell Colonies Generated from MEFs Infected with Lentivirus Containing the KOSM Fusion. Top: Staining of pluripotency markers in induced cell colonies at 200x magnification. Bottom: AP staining at 100x magnification and mor-phology at 40x magnification in induced cell colonies.

Expression of Stem Cell Factors and GFP. Top: Transient ex-pression of KOSM fusion gene in 293T cells confirmed by Western blot. Bottom: GFP fluorescence in MEF cells 3 days after infection with lentivirus containing KOSM fusion.

For efficient packaging of your virus, please see our Lentiviral Packaging Systems on p. 58 and Retroviral Packaging Cell Lines on p. 64 and 67.


Platinum Retroviral Expression Systems for Stem Cells


Platinum ES/EC Retroviral Expression System, Ecotropic 1 kit VPK-303

Platinum ES/EC Retroviral Expression System, Amphotropic 1 kit VPK-304

1 kit VPK-305 Platinum ES/EC Retroviral Expression System, Pantropic

Platinum HSC Retroviral Expression System, Ecotropic 1 kit VPK-306

Platinum HSC Retroviral Expression System, Amphotropic 1 kit VPK-307

Platinum HSC Retroviral Expression System, Pantropic 1 kit VPK-308

34

Retroviral vectors are useful for delivering genes of interest into a host cell where integration into the genome is desired. However, traditional retroviral expression technologies usually result in low viral titers which make gene expression studies difficult.

Higher Viral Yields: Average titer 107 infec-tious units/mL with transient transfection

Versatile: 3 Packaging cell lines for use with nearly any target host species

Optimized for Stem Cell Studies: Specially designed expression systems for either ES/EC cells or hematopoietic stem cells

Amphotropic Ecotropic Pantropic

Human +++ N.S. +++

Mouse +++ +++ +++

Rat +++ +++ +++

Monkey +++ N.S. +++

Cat +++ N.S. +++

Dog +++ N.S. +++

Hamster + N.S. +++

Bird N.S. N.S. +++

Fish N.S. N.S. +++

Frog N.S. N.S. +++

Insect N.S. N.S. +++

Mollusk N.S. N.S. +++

*Virus must be packaged with a pantropic envelope protein such as VSVG. N.S. = Not Suitable

Catalog Number Packaging Cell Line Transfer Vector Envelope Vector Control Vector

VPK-303 Plat-E (Ecotropic) pMCs-Puro ——— pMCs-GFP

VPK-304 Plat-A (Amphotropic) pMCs-Puro ——— pMCs-GFP

VPK-305 Plat-GP (Pantropic) pMCs-Puro pCMV-VSV-G pMCs-GFP

VPK-306 Plat-E (Ecotropic) pMYs-Puro ——— pMYs-GFP

VPK-307 Plat-A (Amphotropic) pMYs-Puro ——— pMYs-GFP

VPK-308 Plat-GP (Pantropic) pMYs-Puro pCMV-VSV-G pMYs-GFP

Suitability of Platinum Retroviral Expression Systems by Host Species.

Components of the Platinum Retroviral Expression Systems for Stem Cells.

Our Platinum Retroviral Expression Systems incorpo-rate superior packaging cell lines and vector technolo-gies to produce high-titer virus with a single plasmid transfection. The Platinum Expression Systems in-clude one of our exclusive Platinum Packaging Cell Lines which already contain the gag and pol genes; the Ecotropic and Amphotropic cells also contain an envelope protein. Simply clone your gene of interest into the vector provided and transfect into the Plati-num cells. If you choose a Pantropic system, simply co-transfect with the VSV-G plasmid provided. The Platinum Expression Systems below are spe-cially designed for superior expression with either ES/EC cells or hematopoietic stem cells. For more infor-mation on our Platinum Expression Systems for a variety of cells, please see page 60.

STEM CELL RESEARCH Retroviral Expression Systems


MEF Feeder Cells


MEF Feeder Cells 5 x 106 cells CBA-310

MEF Feeder Cells, Hygromycin-resistant 5 x 106 cells CBA-313

MEF Feeder Cells, Neomycin-resistant 5 x 106 cells CBA-311

MEF Feeder Cells, Puromycin-resistant 5 x 106 cells CBA-312

35

Our murine embryonic fibroblast (MEF) feeder cells are useful for the maintenance of human or mouse ES cells in their undifferentiated state. Cells must be mitotically inactivated prior to use.

Feeder Cells STEM CELL RESEARCH

SNL 76/7 Passage-Independent Feeder Cells for iPS Culture

The SNL 76/7 is an immortalized cell line derived from mouse fibroblast STO cells which have been transformed with murine LIF and neomycin resistance genes.

Stem Cell Feeders

Leukemia inhibitory factor (LIF) is useful for maintaining the undifferentiated state of mouse embryonic stem (mES) cells. However, LIF does not have the same effect on hu-man embryonic stem (hES) cells. Therefore, hES cells require the use of feeder cells for both derivation and maintenance. We offer a variety of feeder cells for stem cell culture. All feeder cells must be mitotically inactivated prior to use.

Superior Culture: Transformed with LIF gene for better maintenance of undifferentiated state

Versatile: Useful for culture of human and mouse iPS cells and as a feeder for ES cells

Passage-Independent: Immortalized cell line


SNL Feeder Cells 3 x 106 cells CBA-316

Recent Product Citations 1. Osakada, F. et al (2009). In vitro differentiation of retinal cells

from human pluripotent stem cells by small-molecule induction. J. Cell Sci. 132:3169-3179.

2. Liu, Y. et al. (2009). Zeb1 represses Mitf and regulates pigment synthesis, cell proliferation, and epithelial morphology. Invest. Ophthalmol. Vis. Sci. 50:5080-5088.

3. Tsubooka, N. et al. (2009). Roles of Sall4 in the generation of pluripotent stem cells from blastocytes and fibroblasts. Genes Cells 14:683-694.

4. Lan, Z-J. et al. (2009). Extra-germ cell expression of mouse nuclear receptor subfamily 6, group A, member 1 (NR6A1). Biol. Reprod. 80:905-912.

JK1 Passage-Independent Feeder Cells

JK1 is an immortalized CD34+ stromal cell line that supports long-term proliferation of stem cells. It has been shown to maintain capacity for stem cell re-newal even after serial passaging for over one year. JK1 may be used for culture of a variety of cell types including pluripotent ES cells, germ-line derived stem cells such as SPCs and MASCs, and primordial germ cell-derived EG cells.


JK1 Feeder Cells 1 x 106 cells CBA-315

JK1 Cells Support Maintenance of mES Cells. Immunofluores-cence staining of germ cell nu-clear antigen (GCNA). Antibody staining is green; nuclear coun-terstain is blue.


36

CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay

Hematopoietic stem cells (HSCs), when cultured in a suitable semisolid matrix such as methylcellulose supplemented with cytokines & nutrients, proliferate to form discrete cell clusters or colonies. Such HSCs or hematopoietic progenitors are known as colony-forming cells (CFCs). In classic CFC assays, cells are cultured in a 35mm dish for 14-21 days so the colonies can reach a cer-tain size for manual counting, which can be tedious and subjective. The CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay provides a high-throughput method to quantify CFCs in just 7-10 days with no manual cell counting required. Cells are lysed, solu-bilized, and quantified using a fluorescent dye in-cluded in the kit. Alternatively, cells may be recov-ered for further culture and analysis.


CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay Fluorometric 96 Assays CBA-320


Fast Results: 7-10 days vs. 2-3 weeks Plate Reader Convenience: Eliminates manual

counting Easier Reagent Handling: Methylcellulose media

can be handled using a pipet instead of a syringe

Assay Principle for the CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay.

HSC Colony Formation. Human bone marrow derived CD34+ Hematopoietic Progenitor Cells were seeded at 3000 cells/well and cultured for 7-10 days in the absence or presence of growth factors/cytokines. Colonies were quantified according to the assay protocol. A: After 7 days without cytokines. B: After 7 days in presence of cytokines. C: After 10 days in presence of cytokines (hemoglobin visible).

STEM CELL RESEARCH Colony Formation Assays


37

StemTAG™ 96-Well Stem Cell Colony Formation Assay


StemTAG™ 96-Well Stem Cell Colony Formation Assay Fluorometric 96 Assays CBA-325


Our StemTAG™ 96-Well Stem Cell Colony Formation Assay provides a high-throughput method to quantify ES cells in just 7-10 days with no manual cell count-ing required. After colonies are formed, stem cells may be ana-lyzed in 3 ways: 1. Lyse cells, then quantify using a fluorescent dye

included in the kit. 2. Lyse cells, then measure alkaline phosphate ac-

tivity using reagents provided. 3. Recover colonies for further culture and analysis. This assay may be of particular interest for the study of tumor stem cells.

Fast Results: 7-10 days vs. 2-3 weeks using con-ventional methods

Versatile: Quantify cells using fluorescent dye, measure alkaline phosphatase activity, or recover cells for further analysis

Plate Reader Convenience: No manual cell counting required

Anchorage-Independent Growth of Mouse ES-D3 Cells. Top: Phase Contrast. Bottom: Alkaline Phosphatase Staining.

StemTAG™ Stem Cell Colony Formation Assay Principle.

Colony Formation Assays STEM CELL RESEARCH


StemTAG™ PCR Primer Set for Stem Cell Characterization


StemTAG™ PCR Primer Set for Stem Cell Characterization 50 Reactions CBA-303

Total RNA and Protein from Murine ES-D3 Cell Line


Total RNA—Murine Embryonic Stem Cell Line D3 50 µg CBA-304

Total Protein—Murine Embryonic Stem Cell Line D3 500 µg CBA-305

38

Pluripotent stem cells can differentiate into cells derived from all three embryonic germ layers: endoderm, mesoderm and ectoderm. Our StemTAG™ PCR Primer Set provides an efficient system for monitoring ES cell differentiation/undifferentiation. Seven primer sets are included: primers for two widely studied stem cell mark-ers (Oct-4 and NANOG), one marker for each embryonic germ layer (AFP/Endoderm, Flk-1/Mesoderm and NCAM/Ectoderm), and two controls (GAPDH and ß-Actin). Primers are suitable for either end-point or real-time (quantitative) PCR.

STEM CELL RESEARCH Alk Phos Assays, Primers, RNA/Protein

StemTAG™ Alkaline Phosphatase Kits

StemTAG™ Alkaline Phosphatase Staining Kit. Murine embry-onic stem cells (ES-D3) were maintained in an undifferentiated state with LIF. To induce differentiation, LIF was withdrawn over several days. Various differentiation events were observed: cells became flattened and enlarged with reduced proliferation. On day 5, cells were stained according to the assay protocol.


StemTAG™ Alkaline Phosphatase Staining and Activity Assay Kit ICC & Colorimetric 2 x 100 Assays CBA-302

ICC & Fluorometric 2 x 100 Assays CBA-308



StemTAG™ Alkaline Phosphatase Staining Kit (Red) ICC 100 Assays CBA-300

StemTAG™ Alkaline Phosphatase Activity Assay Kit

StemTAG™ Alkaline Phosphatase Staining Kit (Purple) ICC 100 Assays CBA-306

The StemTAG™ Alkaline Phosphatase Staining and Activity Assay Kits monitor AP activity via both immu-nocytochemistry staining and a colorimetric 96-well plate-based activity assay. The staining and activity assay kits are also sold separately.

Fast Results: Staining and Activity Assay proto-cols each take less than 1 hour

Versatile: Useful for human ES, EG and EC cells, as well as mouse ES and EG cells

Recent Product Citations 1. Lee, J. et al. (2010). Ultraviolet A regulates adipogenic differen-

tiation of human adipose tissue-derived mesenchymal stem cells via up-regulation of kruppel-like factor 2. J. Biol. Chem. 285:32647-32656. (CBA-300)

2. Izadyar, F. et al. (2008). Generation of multipotent cell lines from a distinct population of male germ line stem cells. Reproduction 135:771-784. (CBA-300)

3. Kim, Y. et al. (2009). Cyclin-dependent kinase 2-associating protein 1 commits murine embryonic stem cell differentiation through retinoblastoma protein regulation. J. Biol. Chem. 284:23405-23414. (CBA-301)

4. Yue, Y. et al. (2008). A single intravenous injection of adeno-associated virus serotype-9 leads to whole body skeletal muscle transduction in dogs. Mol. Ther. 16(12):1944-1952. (CBA-301)

5. Ghosh, A. et al (2007). Efficient whole-body transduction with trans-splicing AAV vectors. Mol. Ther. 15(4):750-755. (CBA-301)


VIRAL EXPRESSION Viral Vector Overview

Recombinant Viral Gene Delivery

Recombinant viral vectors provide a powerful means of delivering a gene into a target cell. There are many viral vectors available, and there are pros and cons to each. Use the fol-lowing table to select the best viral vector for your research.

Comparison of Viral Vectors for Gene Delivery Adeno-Associated

Virus (AAV) (p. 41-48)

Adenovirus (p. 49-56)

Lentivirus (HIV-1, FIV, SIV)

(p. 57-63)

Retrovirus (MMLV)

(p. 64-72)

Gene Expression Transient or Stable

Transient Transient or Stable

Stable

Will Infect Dividing Cells Yes Yes Yes Yes

Will Infect Non-Dividing Cells Yes Yes Yes No

Integrates into Target Cell Genome No* No Yes Yes

Relative Viral Titer XXX XXXX XXX XX

Relative Transduction Efficiency XXX XXXX XXX XX

Immune Response in Target Cells Very Low High Low Moderate

40

Typical Workflow for Viral Gene Delivery

Clone Gene of Interest

Package Virus

Measure Titer

Concentrate & Purify

Infect Target Cell

Viral Expression

Plasmid

Packaging Plasmids and Cells

Purification & Concentration

Kit

Viral Transduction

Reagents

Viral Quantitation

Kit


*Native AAV can integrate, but recombinant AAV rarely does.

Cell Biolabs offers kits and reagents for every step in your workflow.

VIRAL EXPRESSION AAV Expression

41

Adeno-Associated Virus Kits & Reagents

Adeno-associated virus (AAV) is less immunogenic than adenovirus or retrovirus. We offer a comprehensive line of AAV kits and reagents to ensure you get the best expression from your AAV expression studies:

AAV Helper Free Systems

AAV Helper Free Systems are available for a variety of formats and

serotypes: AAV Complete Expression Systems

contain all packaging plasmids plus an expression vector and a GFP control vector: p. 42-45

AAV Packaging Systems contain the pHelper plasmid and a serotype-specific Rep-Cap plasmid for use with your own expression construct: p. 45

If you have an AAV packaging system for one serotype and want to try another, choose one of 8 different AAV Rep-Cap Plasmids from native serotypes 1 through 6 plus AAV-DJ and AAV-DJ/8: p. 45

If you already have an AAV packaging system and need a cloning vector, choose one of 10 different AAV Expres-sion Vectors available individually: p. 46

Want to make a control virus? Choose one of our AAV Control Vectors: p. 46

Production of recombinant AAV requires certain genes from the adenovirus genome, which means that an adenovirus usually needs to be present. The AAV Helper Free System eliminates the need for a helper adenovirus. Most of the required adenoviral genes (E2A, E4 and VA RNA) are provided in a pHelper plasmid, while the required E1 gene is pro-vided by the 293 packaging cells.

Safer: pHelper plasmid eliminates the need for a helper virus

Flexible: Packaging vectors and expression vec-tors available separately or as one complete sys-tem, so you only order what you need

Expandable: All plasmids are provided individually, not in a mixture, so you can amplify in competent cells

Premade AAV Controls Purification Kits Quantitation / Titer Kit Transduction Kits

Helper Free Expression Systems Helper Free Packaging Systems Expression & Control Vectors Viral Packaging Cell Line

Gene Delivery using the AAV Helper Free System.





AAV-DJ Helper Free Expression System 1 kit VPK-410-DJ

AAV-DJ Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-DJ

AAV-DJ Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-DJ

AAV-DJ Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-DJ

AAV-DJ Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-DJ

AAV-DJ Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-DJ

AAV-DJ Helper Free Promoterless Expression System 1 kit VPK-411-DJ

AAV-DJ Helper Free shRNA Expression System (Puro) 1 kit VPK-412-DJ

AAV-DJ/Helper Free shRNA Expression System (GFP) 1 kit VPK-413-DJ

scAAV-DJ Helper Free Expression System 1 kit VPK-430-DJ

AAV Helper Free Complete Expression Systems

AAV Helper Free Complete Expression Systems con-tain everything you need to produce high-titer recom-binant adeno-associated virus:

pHelper Plasmid Rep-Cap Plasmid (serotype specific) GFP Control Vector Choice of 10 AAV Expression Vectors:

Gene Expression (CMV or no promoter) shRNA (U6 promoter) Self complementary (scAAV)

AAV Helper Free Expression Systems are available for the following serotypes: Native serotypes 1-6 AAV-DJ, engineered by DNA family shuffling to

form a hybrid capsid from 8 different native sero-types; provides significantly higher infectivity rates in vitro (see table below)

AAV-DJ/8, a mutant of AAV-DJ that exhibits in-creased uptake in brain and other tissues in vivo, similar to serotypes 8 and 9

Relative Infectivity Rates of AAV Serotypes. Normalized to AAV-2 = 100. ND = Not determined.

Cell Line Cell or Tissue Source AAV-1 AAV-2 AAV-3 AAV-4 AAV-5 AAV-6 AAV-8 AAV-9 AAV-DJ

AAV-DJ/8

Huh-7 Hu Liver 13 100 2.5 0.0 0.1 10 0.7 0.0 500 0.2

HEK293 Hu Kidney 25 100 2.5 0.1 0.1 5 0.7 0.1 500 0.3

HeLa Hu Cervix 3 100 2.0 0.1 3.7 1.0 0.2 0.1 667 0.2

HepG2 Hu Liver 3 100 16.7 0.3 1.7 5 0.3 ND 1250 0.5

Hep1A Ms Liver 20 100 0.2 1.0 0.1 1.0 0.2 0.0 400 0.1

911 Hu Retina 17 100 11.1 0.2 0.1 17 0.1 ND 500 0.0

CHO Hm Ovary 100 100 14.3 1.4 333 50 10.0 1.0 25000 5.0

COS Si Kidney 33 100 33 3.3 5.0 14 2.0 0.5 500 0.3

MeWo Hu Skin 10 100 20 0.3 6.7 10 1.0 0.2 2857 1.0

NIH3T3 Ms Fibroblasts 10 100 2.9 2.9 0.3 10 0.3 ND 500 0.1

A549 Hu Lung 14 100 20 ND 0.5 10 0.5 0.1 1000 0.1

HT1180 Hu Fibroblasts 20 100 10.0 0.1 0.3 33 0.5 0.1 333 0.2

Monocytes Hu Primary Monocytes 1111 100 ND ND 125 1429 ND ND 100 ND

Immature DC Hu Monocyte-derived DC 2500 100 ND ND 222 2857 ND ND 200 ND

Mature DC Hu Monocyte-derived DC 2222 100 ND ND 333 3333 ND ND 100 ND

AAV-DJ Helper Free Complete Expression Systems

AAV-DJ/8 Helper Free Complete Expression Systems



AAV-1 Helper Free Complete Expression Systems


AAV-DJ/8 Helper Free Expression System 1 kit VPK-410-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-DJ-8

AAV-DJ/8 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-DJ-8

AAV-DJ/8 Helper Free Promoterless Expression System 1 kit VPK-411-DJ-8

AAV-DJ/8 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-DJ-8

AAV-DJ/8 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-DJ-8

scAAV-DJ/8 Helper Free Expression System 1 kit VPK-430-DJ-8


AAV-1 Helper Free Expression System 1 kit VPK-410-SER1

AAV-1 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-SER1

AAV-1 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER1

AAV-1 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER1

AAV-1 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-SER1

AAV-1 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-SER1

AAV-1 Helper Free Promoterless Expression System 1 kit VPK-411-SER1

AAV-1 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-SER1

AAV-1 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-SER1

scAAV-1 Helper Free Expression System 1 kit VPK-430-SER1



























AAV Helper Free Packaging Systems

AAV Helper Free Packaging Systems contain everything found in the Complete Expression Systems, with the exception of the AAV expression vector. This is an ideal choice if you already have an AAV construct containing your gene of interest. All plasmids are provided individually, not as a packaging mixture.


AAV-2 Helper Free Packaging System 1 kit VPK-402




AAV-DJ Helper Free Packaging System 1 kit VPK-400-DJ

AAV-DJ/8 Helper Free Packaging System 1 kit VPK-400-DJ-8



AAV Rep-Cap Plasmids (Serotype-Specific)

AAV Rep-Cap plasmids allow you to make recombinant AAV of a specific serotype. These plasmids are ideal if you already have an AAV packaging system for a different serotype. Just substitute one of these plasmids into your AAV Helper Free Packaging System or Expression System.

Product Name Catalog Number

pAAV-DJ Vector VPK-420-DJ

pAAV-DJ/8 Vector VPK-420-DJ-8

pAAV-RC1 Vector VPK-421



pAAV-DJ Vector VPK-420-DJ

pAAV-DJ/8 Vector VPK-420-DJ-8




46

AAV Expression Vectors


pAAV-MCS Expression Vector 10 µg VPK-410

pAAV-IRES-Puro Expression Vector 10 µg VPK-415

pAAV-IRES-Neo Expression Vector 10 µg VPK-416

pAAV-IRES-Hygro Expression Vector 10 µg VPK-417

pAAV-IRES-GFP Expression Vector 10 µg VPK-418

pAAV-IRES-Bsd Expression Vector 10 µg VPK-419

pAAV-MCS Promoterless Expression Vector 10 µg VPK-411

pAAV-U6-Puro Expression Vector 10 µg VPK-412

pAAV-U6-GFP Expression Vector 10 µg VPK-413

Cloning Capacity

3 kb

1.8 kb

1.6 kb

1.4 kb

1.7 kb

2 kb

3.9 kb

2.2 kb

2.1 kb

pscAAV-MCS Expression Vector 1.5 kb 10 µg VPK-430

AAV Control Plasmids


pAAV-GFP Control Vector 10 µg AAV-400

pAAV-Cre Control Vector 10 µg AAV-401

pAAV-LacZ Control Vector 10 µg AAV-402

pscAAV-GFP Control Vector 10 µg AAV-410


AAV Premade Control Viruses


AAV1-GFP Control Virus 50 µL AAV-301

AAV2 Null Control Virus 50 µL AAV-300

AAV2-Cre Control Virus 50 µL AAV-310


AAV2-Luc Control Virus 50 µL AAV-320




All AAV premade viruses are provided at a concentration of 1 x 1012 GC/mL.

Each of our AAV Expression Vectors may be used with any of our AAV Helper Free Systems, regardless of AAV serotype. Choose one of these vectors when you already have an AAV Packaging System but may want to use a different promoter or selection marker.

Choose one of our AAV control vectors when you already have an AAV Packaging System and want to make a transduction control virus.

Recent Product Citation Sen, Y. et al. (2014). TDP-43 causes differential pathology in neu-ronal versus glial cells in the mouse brain. Human Mol. Genet. 10.1093/hmg/ddt662. (VPK-410)


47

ViraBind™ AAV Purification Kits

Purification of AAV via ultracentrifugation can be tedious and time-consuming, and may result in low yields. ViraBind™ AAV Purification Kits use a one-step proprietary matrix followed by further purification and concen-tration using a centrifugal concentrator. The result is a higher AAV yield with high purity in a fraction of the time. Kits are suitable for AAV-2 or AAV-DJ; they will not work with other AAV serotypes.

High Purity: No contamination bands as seen on SDS gel

Fast Results: Obtain purified virus in about 3 hours

High Yields: Recovery rate >60%

Electrophoretic Profile of Purified AAV2-GFP.

Product Name Capacity/Prep Size Catalog Number

ViraBind™ AAV Purification Kit Two 10-cm dishes 10 Preps VPK-140

ViraBind™ AAV Purification Mega Kit 2 Preps VPK-141

10 Preps VPK-141-5 Ten 15-cm dishes

Purification Procedure for the ViraBind™ AAV Purification Kit.

293AAV Cell Line


1 x 106 cells AAV-100 293AAV Cell Line

Our 293AAV cell line is selected from the parental 293 cell line for larger surface area, flattened morphology, and firmer attachment to culture plates, resulting in production of higher yields of AAV.


Recent Product Citation Uchida, S. et al. (2010). Early life stress enhances behavioral vulnerability to stress through the activation of REST4-mediated gene transcription in the medial prefrontal cortex of rodents. J. Neurosci. 30:15007-15018. (VPK-140)


QuickTiter™ AAV Quantitation Kit


QuickTiter™ AAV Quantitaiton Kit Fluorometric 20 Assays VPK-145

Fast Results: Obtain purified virus in less than 2 hours

High Sensitivity: Limit of detection 1 x 109 GC/mL from unpurified supernatant or 5 x 1010 GC/mL from purified AAV

Traditional AAV Quantitation by dot blot can be tedi-ous, time consuming, and suffer from high inter-assay variability. Our QuickTiter™ AAV Quantitation Kit uses a proprietary technology to quantify AAV nucleic acid content of unpurified AAV-2 or AAV-DJ, or from purified AAV of any serotype.

0

25

50

75

100

125

150

175

200

0 25 50 75 100 125 150

AAV DNA STD (ng)

RF

U

0

5

10

15

20

0 2 4 6 8 10

AAV DNA STD (ng)

RF

U

AAV-2 DNA Standard Curve. The QuickTiter™ AAV-2 DNA Standard was diluted as described in the assay protocol. Fluorescence was measured on a SpectraMax Gemini XS Fluorometer (Molecular Devices) with a 485/538 nm filter set and 530 nm cutoff.

ViraDuctin™ AAV Transduction Reagent

Product Name Size* Catalog Number

10 Transductions AAV-200

50 Transductions AAV-201 ViraDuctin™ AAV Transduction Reagent

48

*Number of transductions performed in 35mm culture dishes. May be modified for use in culture plates or larger dishes. See product insert.

Our ViraDuctin™ AAV Transduction Reagent can significantly increase the transduction efficiency of AAV vectors in both dividing and non-dividing cells. Increases are greatest in non-dividing cells, but even cells in S-phase show a noticeable increase in trans-duction efficiencies.

Higher Efficiencies: Significantly increase rate of infection of host cells

Low Toxicity: No noticeable effect on cell viability Universal: Suitable for use with both dividing and

non-dividing cells

Successful gene expression studies using AAV depend on high transduction efficiencies into host cells. Infection rates appear to be highest in S-phase cells, which can account for a very small fraction of a cell population.


VIRAL EXPRESSION Adenoviral Expression

49

Adenoviral Expression Kits & Reagents

Recombinant adenoviruses are excellent tools for introducing genetic material into host cells, since they can infect a variety of mammalian cell types with high efficiency. They re-main epichromosal upon infection, so they are only suitable for transient gene expression. We offer a complete workflow solution to your adenoviral expression studies:

Purification Kits Quantitation / Titer Kits Transduction Reagents

Viral Expression Systems Viral Packaging Cell Line Premade Recombinant Adenoviruses

RAPAd® Adenoviral Expression Systems

Adenovirus Production using the RAPAd® Adenoviral Expression System.

Compared to other adenoviral expression systems, RAPAd® Adenoviral Expression Systems produce recombinant adenovirus in a much shorter time (about 2-3 weeks) with a substantial reduction in wild-type adenovirus. The RAPAd systems use a backbone vector from which the 5’ ITR, packaging signal and E1 sequences have been removed. Addi-tionally, serial amplification of the recombinant ade-novirus does not increase the level of replication-competent adenovirus.

Virtually No Wild-Type Virus: Backbone vector engineered to produce <300 wild-type plaques per 109 particles, compared with 104-106 WT plaques per 109 VP with most other methods

Faster Production: Virus generated in 2-3 weeks compared to a few months with traditional methods

7 Complete Systems: Choose CMV or RSV for gene expression, EF-1 for miRNA expression, U6 for shRNA, or clone your own promoter along with your gene of interest using our Universal system

Product Name Promoter Size Catalog Number

RAPAd® Universal Adenoviral Expression System None 1 Kit VPK-250

RAPAd® RSV Adenoviral Expression System RSV 1 Kit VPK-251

RAPAd® CMV Adenoviral Expression System CMV 1 Kit VPK-252

RAPAd® miRNA Adenoviral Expression System EF-1 1 Kit VPK-253

RAPAd® Bicistronic Adenoviral Expression System (GFP) CMV 1 Kit VPK-254

RAPAd® shRNA Adenoviral Expression System (Puro) U6 1 Kit VPK-255

RAPAd® shRNA Adenoviral Expression System (GFP) U6 1 Kit VPK-256


Recent Product Citations 1. Kothari, H. et al. (2010). Cystine 186-cystine 209 disulfide bond

is not essential for the procoagulant activity of tissue factor or for its de-encryption. Blood 115:4273-4283. (VPK-252)

2. Li, P. et al. (2013). MicroRNA-638 is highly expressed in hu-man vascular smooth muscle cells and inhibits PDGF-BB-induced cell proliferation and migration through targeting or-phan nuclear receptor NOR1. Cardiovasc. Res. 10.1093/cvr/cvt082. (VPK-253)

50


Don’t have time to make your own adenovirus? Are you studying the expression of multiple genes? Rely on our premade recombinant adenoviruses that al-ready contain a gene of interest. All of Cell Biolabs’ premade recombinant adenoviruses are provided as 50 µl aliquots at a concentration of 1 x 1011 viral parti-cles/mL in TBS with 10% glycerol.

Target Name Catalog Number

Null Control (No gene) ADV-001

-Galactosidase ADV-002

Cre ADV-005

Firefly Luciferase ADV-008

GFP ADV-004

SEAP (Secretory Alkaline Phosphatase) ADV-003

Controls and Reporter Genes

Recent Product Citations 1. Matsushita, T. et al. (2009). FGFR3 promotes synchondosis

closure and fusion of ossification centers through the MAPK pathway. Hum. Mol. Genet. 18:227-240. (ADV-001)

2. Zhang, Z. et al (2013). MEK inhibition leads to lysosome-mediated Na+/I– symporter protein degradation in human breast cancer cells. Endocr. Relat. Cancer 20:241-250. (ADV-002)

3. Schramm, C. et al (2012). The PTPN11 loss-of-function muta-tion Q510E-Shp2 causes hypertrophic cardiomyopathy by dys-regulating mTOR signaling. Am. J. Physiol. Heart Circ. Physiol. 302:H231-H243. (ADV-002)

4. Salvati, E. et al. (2014). Evidence for G-quadruplex in the pro-moter of vegfr-2 and its targeting to inhibit tumor angiogenesis. Nucleic Acids Res. 42:2945-2957. (ADV-004)

5. Lu, D. et al. (2012). Peroxisome proliferator-activated receptor-coactivator-1alpha enhances engraftment and angiogenesis of mesenchymal stem cells in diabetic hindlimb ischemia. Diabe-tes 61:1153-1159. (ADV-004)

6. Kato, H. et al. (2011). Wnt/ß-Catenin pathway in podocytes integrates cell adhesion, differentiation and survival. J. Biol. Chem. 286:26003-26015. (ADV-005)

Premade Recombinant Adenoviruses

Cancer/Tumor Antigens


Carbonic Anhydrase 9 (CA9) ADV-602

Carcinoembryonic Antigen (CEA) ADV-604

NY-ESO-1 ADV-601

Blood Vessel Formation After 3 Days. Purified Ad-Null or Ad-VEGF viruses were applied to a 10-day old CAM (chick chorioal-lanoic membrane). Results were visualized by stereomicroscope.

Angiogenesis


VEGF ADV-101

HIF-1 ADV-100

Recent Product Citations 1. Kelber, J.A. et al. (2012). KRas induces a Src/PEAK1/ErbB2

kinase amplification loop that drives metastatic growth and therapy resistance in pancreatic cancer. Cancer Res. 72:2554-2564. (ADV-101)

2. Qiu, X. et al. (2012). Combined strategy of mesenchymal stem cell injection with vascular endothelial growth factor gene ther-apy for the treatment of diabetes-associated erectile dysfunc-tion. J. Androl. 33:37-44. (ADV-101)

3. Stoletov, K. et al. (2010). Visualizing extravasation dynamics of metastatic tumor cells. J. Cell Sci. 123:2332-2341. (ADV-101)

4. Serban, D. et al. (2008). H-ras regulates angiogenesis and vascular permeability by activation of distinct downstream ef-fectors. Circ. Res. 102(11):1350-1358. (ADV-101)

293AD Cell Line for Adenoviral Packaging and Amplification

The 293AD cell line is derived from the parental 293 cell line, but has been specifically selected for adeno-virus applications and offers advantages over conven-tional 293 cells: flattened morphology, firm attach-ment to culture plates, and a larger surface area for superior transfection and greater viral yields.

Recent Product Citations 1. Peng, D. et al. (2011). Glutathione peroxidase 7 protects

against oxidative DNA damage in oesophageal cells. Gut 61:1250-1260. (AD-100)

2. Kothari, H. et al. (2010). Cystine 186-cystine 209 disulfide bond is not essential for the procoagulant activity of tissue factor or for its de-encryption. Blood 115:4273-4283. (AD-100)


293AD Cell Line 1 x 106 Cells AD-100



51

Premade Recombinant Adenoviruses, continued

Actin Cytoskeleton Staining. Cos-7 cells were infected with puri-fied Ad-Ras V12 (ADV-146) at 50 MOI (multiplicity of infection). Membrane ruffling was visualized by staining the actin cytoskeleton with Rhodamine-coupled Phalloidin.

Cytoskeleton Regulation / Small GTPase


Cdc42 ADV-152

Cdc42 L61 (Constitutively Active) ADV-154

Cdc42 N17 (Dominant Negative) ADV-153

PAK1 ADV-202

PAK1 (H83L, H86L) ADV-203

PAK1 (H83L, H86L, K299R) ADV-205

PAK1 (K299R) ADV-207

PAK1 (L107E, T423E) ADV-206

PAK1 (T423E) ADV-204

PAK1 (Kinase Domain) ADV-209

PAK1 (Regulatory Domain) ADV-208

Rac1 ADV-149

Rac1 L61 (Constitutively Active) ADV-151

Rac1 N17 (Dominant Negative) ADV-150

Recent Product Citations 1. Black, S.A. et al. (2008). TGFß1 stimulates connective tissue

growth factor (CCN2/CTGF) expression in human gingival fibroblasts through a RhoA-independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGFß1-induced CCN2/CTGF expression. J. Biol. Chem. 283:10835-10847. (ADV-145, ADV-150, ADV-153, ADV-156)

2. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires vß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Cell Biol. 23:1104-1114. (ADV-150)

3. Thomas, M.A. et al. (2009). E4orf1 limits the oncolytic potential of the E1B-55K-deleted adenovirus. J. Virol. 83:2406-2416. (ADV-150)

4. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150, ADV-153, ADV-154)

5. Salvati, E. et al. (2014). Evidence for G-quadruplex in the pro-moter of vegfr-2 and its targeting to inhibit tumor angiogenesis. Nucleic Acids Res. 42:2945-2957. (ADV-151, ADV-157)

6. Cheng, Z.-J. et al. (2010). Co-regulation of caveolar and Cdc42-dependent fluid phase endocytosis by phosphocaveolin-1. J. Biol. Chem. 285:15119-15125. (ADV-153)

7. Neal M. et al. (2013). A critical role for TLR4 induction of auto-phagy in the regulation of enterocyte migration and the patho-genesis of necrotizing enterocolitis. J. Immunol. 190:3541-3551. (ADV-156, ADV-157)

8. Nie, J. et al. (2013). SAD-A kinase controls islet ß-cell size and function as a mediator of mTORC1 signaling. PNAS 110:13857-13862. (ADV-204, ADV-207)

9. Nie, J. et al. (2013). Synapses of amphids defective (SAD-A) kinase promotes glucose-stimulated insulin secretion through activation of p21-activated kinase (PAK1) in pancreatic ß-cells. J. Biol. Chem. 287:26435-26444. (ADV-204, ADV-207)

Cell Cycle & Transcription Regulation


p53 ADV-501

p53 (Temperature Sensitive Mutant) ADV-502

p68 RNA Helicase ADV-505

Myogenin ADV-509


DCC ADV-504

MyoD ADV-508


Ras N17 (Dominant Negative) ADV-145

Ras V12 (Constitutively Active) ADV-146

Ras V12C40 ADV-148

Ras V12S35 ADV-147

Rho L63 (Constitutively Active) ADV-157

Rho N19 (Dominant Negative) ADV-156

SDF-1 ADV-210


Recent Product Citation Nguyen, N. et al. (2014). Mitsugumin 53 (MG53) ligase ubiquiti-nates focal adhesion kinase during skeletal myogenesis. J. Biol. Chem. 289:3209-3216. (ADV-508)

52


MAP Kinase Signaling



Cdc42 ADV-152



ERK2 ADV-112

ERK2 (Dominant Negative) ADV-113

ERK5 (BMK1) ADV-116

ERK5 (Dominant Negative) ADV-117

Interferon- ADV-103

Interleukin-2 ADV-102

JNK1 ADV-114

JNK1 (Dominant Negative) ADV-115

MAPKAPK2 ADV-137

MAPKAPK2 (Dominant Negative) ADV-138

MAPKAPK2 (Constitutively Active) ADV-139

MEK1 (Dominant Negative) ADV-118

MEK1 (Constitutively Active) ADV-119

MEK5 ADV-129

MEK5 (Dominant Negative) ADV-130

MEK5 (Constitutively Active) ADV-131

MEKK1 ADV-135

MEKK1 (Dominant Negative) ADV-136

MEKK3 ADV-162

Immunoblot of Various Cell Signaling Targets. HUVEC cells were in-fected with purified Ad-Null (ADV-001), Ad-Ras V12 (ADV-146), Ad-Ras V12S35 (ADV-147), Ad-Ras V12C40 (ADV-148), Ad-MEK1 (ADV-119), and Ad-Rac1 L61 (ADV-151) at 10 MOI (multiplicity of infection). Cell lysates were ana-lyzed for gene expres-sion and ERK activation.


Rac1 ADV-149



Raf1 ADV-132

Raf1 (Dominant Negative) ADV-133

Raf1 (Constitutively Active) ADV-134





SOK ADV-142

SOK (Dominant Negative) ADV-143

SOK (Constitutively Active) ADV-144

Tac-Rac1 (Membrane Targeting) ADV-164

MKK6 (Dominant Negative) ADV-124

MKK6 (Constitutively Active) ADV-125

MKK7 ADV-126



myr-Rac1 ADV-163

p38 ADV-104

p38 (Dominant Negative) ADV-105

p38 ADV-106


p38 ADV-108



PRAK (Dominant Negative) ADV-141





MKK6 ADV-123

MKK3 ADV-120

See following page for recent citations using these adenoviruses


53



Tyrosine Kinases and PKCs


CSK ADV-405

CSK (Dominant Negative) ADV-406

Fyn ADV-403

Fyn (Dominant Negative) ADV-404

PKC- (Dominant Negative) ADV-410



shAkt1 ADV-417

shAkt2 ADV-418

Src ADV-401

Recent Product Citations 1. Harbrecht, B.G. et al. (2012). Insulin inhibits hepatocyte iNOS

expression induced by cytokines by an Akt-dependent mecha-nism. Am. J. Physiol. Gastrointest. Liver Physiol 302:G116-G122. (ADV-105)

2. Jones, S.W. et al. (2009). Mitogen-activated protein kinase-activated protein kinase (MK2) modulates key biological path-ways associated with OA disease pathology. Osteoarthritis and Cartilage 17:124-131. (ADV-105)

3. Kim, J.M. et al. (2008). Inhibition of apoptosis in Bacteroids fragilis enterotoxin-stimulated intestinal epithelial cells through the induction of c-IAP-2. Eur. J. Immunol. 38(8):2190-2199. (ADV-105)

4. Lee, J.Y. et al. (2007). Effects of transcription factor activator protein-1 on interleukin-8 expression and enteritis in response to Clostridium difficile toxin A. J. Mol. Med. 85:1393-1404. (ADV-105, ADV-115)

5. Monick, M. et al. (2008). Constitutive ERK MAPK activity regu-lates macrophage ATP production and mitochondrial integrity. J. Immunol. 180:7485-7496. (ADV-112, ADV-113, ADV-118, ADV-119)

6. Samuel, I. et al. (2008). Enteral exclusion increases MAP kinase activation and cytokine production in a model of gall-stone pancreatitis. Pancreatology 8(1):6-14. (ADV-113)

7. Wang, X. et al. (2007). Human immunodeficiency virus prote-ase inhibitor ritonavir inhibits cholesterol efflux from human macrophage-derived foam cells. Am. J. of Pathology 171:304-314. (ADV-113)

8. Jiang, S. et al. (2011). Role of inhibitory kB kinase and c-Jun NH2-terminal kinase in the development of hepatic insulin re-sistance in critical illness diabetes. Am. J. Physiol. Gastrointest. Liver Physiol 301:G454-G463. (ADV-115)

9. Zhang, Z. et al. (2013). MEK inhibition leads to lysosome-mediated Na+/I– symporter protein degradation in human breast cancer cells. Endocr. Relat. Cancer 20:241-250. (ADV-118)

10. Matsushita, T. et al. (2009). FGFR3 promotes synchondosis closure and fusion of ossification centers through the MAPK pathway. Hum. Mol. Genet. 18:227-240. (ADV-119)

11. Yoon, C-H. et al. (2009). Activation of p38 mitogen-activated protein kinase is required for death receptor-independent cas-pase-8 activation and cell death in response to sphingosine. Mol. Cancer Res. 7(3):361-370. (ADV-119)

12. Tan, S.H. et al. (2009). Regulation of cell proliferation and mi-gration by TAK1 via transcriptional control of von Hippel-Lindau tumor suppressor. J. Biol. Chem. 284:18047-18058. (ADV-128)

13. Wu, Y. et al. (2012). ERK5 regulates glucose-induced in-creased fibronectin production in the endothelial cells and in the retina in diabetes. Invest. Ophthalmol. Vis. Sci. 53:8405-8413. (ADV-130)

14. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires vß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Cell Biol. 23:1104-1114. (ADV-150)

15. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150)

16. Neal, M. et al. (2013). A critical role for TLR4 induction of auto-phagy in the regulation of enterocyte migration and the patho-genesis of necrotizing enterocolitis. J. Immunol. 190:3541-3551. (ADV-156, ADV-157)

17. Taniguchi, C. et al. (2007). The p85a regulatory subunit of phosphoinositide 3-kinase potentiates c-Jun N-terminal kinase-mediated insulin resistance. Mol. Cell Biol. 27:2830-2840. (ADV-161)

MAP Kinase Signaling, continued NFB Signaling


IB- ADV-301

IB- S32A (Dominant Negative) ADV-302

IKK- ADV-305

IKK- (Dominant Negative) ADV-303

NOD2 ADV-308

Rel B ADV-304

Recent Product Citations 1. Johnston, R.K. et al. (2009). ß3-integrin mediated ubiquitination

activates survival signaling during myocardial hypertrophy. FASEB J. 23(8):2759-2771. (ADV-302)

2. Martin, A.P. et al. (2008). Lapatinib resistance in HCT116 cells is mediated by elevated MCL-1 expression and decreased BAK activation and not by ERBB receptor kinase mutation. Mol. Pharmacol. 74:807-822. (ADV-302)

3. Richardson, W.M. et al. (2010). Nucleotide-binding oligomeri-zation domain-2 inhibits toll-like receptor-4 signaling in the intestinal epithelium. Gastroenterology 139(3):904-917. (ADV-308, ADV-309)

Recent Product Citations 1. Koh, W. et al. (2009). Formation of endothelial lumens requires a

PKC-, Src-, Pak-, and Raf-kinase dependent signaling cascade downstream of Cdc42 activation. J. Cell Sci. 122:1812-1822. (ADV-401, ADV-405, ADV-406)

2. Lecuona, E. et al. (2009). Ubiquitination participates in the ly-sosomal degradation of the Na,K-ATPase in steady state condi-tions. Am. J. Respir. Cell Mol. Biol. 41(6):617-679. (ADV-412)

3. Vadasz, I. et al. (2008). AMP-activated protein kinase regulates CO2-induced alveolar epithelial dysfunction in rats and human cells by promoting Na,K-ATPase endocytosis. J. Clin. Invest. 118(2):752-762. (ADV-412)

4. Briva, A. et al. (2007). High CO2 levels impair alveolar epithelial function independent of pH. PLoS ONE 2(11):e1238. (ADV-412)



54

ViraBind™ Adenovirus Purification Kits

Purification of Recombinant Ad--Gal. Ad-β-Gal was purified according to the assay protocol. Each purification fraction was used to infect A549 cells in a 12-well plate. After 48 hr, cells were scored using our β-Galactosidase Staining Kit (p. 92).


ViraBind™ Adenovirus Miniprep Kit 1 x 1011 VP 10 Preps VPK-099

ViraBind™ Adenovirus Purification Kit 2.5 x 1012 VP 10 Preps VPK-100

Purification of viruses via cesium chloride (CsCl) ul-tracentrifugation procedures can be tedious and time-consuming. ViraBind™ Adenovirus Purification Kits use an efficient system for quick adenoviral purifica-tion with high recovery. No ultracentrifugation is re-quired. Kits use either a spin column or syringe filter for high purity adenovirus (see selection guide).

High Viral Yield: >90% recovery High Quality: Provides quality of CsCl procedures,

but in much less time Faster Results: 30 minutes (1-2 hrs for Mega kit) User-Friendly Protocol: No gradient preparation

or ultracentrifugation steps

0

20

40

60

80

100

Viral S

upern

atan

t

Flow-th

rough

1st W

ash

2nd

Was

h

Elutio

n

Re

lati

ve

VP

(%

)

Recent Product Citations 1. Wilkins, H. et al. (2013). Mitochondrial glutathione transport is a

key determinant of neuronal susceptibility to oxidative and nitrosative stress. J. Biol. Chem. 288:5091-5101. (VPK-099)

2. Wang, Y.S. et al. (2012). MicroRNA-195 regulates vascular smooth muscle cell phenotype and prevents neointimal forma-tion. Cardiovasc. Res. 95:517-526. (VPK-099)

3. Kirui, J.K. et al. (2010). Gßgamma signaling promotes breast cancer cell migration and invasion. J. Pharmacol. Exp. Ther. 333:393-403. (VPK-099)

4. Scallan, C. et al. (2013). An adenovirus-based vaccine with a double-stranded RNA adjuvant protects mice and ferrets against H5N1 avian influenza in oral delivery models. Clin. Vaccine Immunol. 20:85-94. (VPK-100)

5. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phos-phorylation of vascular endothelial cadherin induced by inva-sive breast cancer cells. J. Biol. Chem. 287:32981-32992. (VPK-100)

6. Chen, F. et al. (2011). Dynamic regulation of PDX-1 and FoxO1 expression by FoxA2 in dexamethasone-induced pan-creatic ß-cells dysfunction. Endocrinology 152:1779-1788. (VPK-100)

7. Prasad, S.S. et al. (2011). Enzymatic activities of the human AGPAT isoform 3 and isoform 5: localization of AGPAT5 to mitochondria. J. Lipid Res. 52:451-462. (VPK-100)

8. Agarwal, A.K. et al. (2010). Enyzmatic activity of the human 1-acylglycerol-3-phosphate-O-acyltransferase isoform 11: upregulated in breast and cervical cancers. J. Lipid Res. 51:2143-2152. (VPK-100)

9. Triulzi, C. et al. (2010). Antibody-dependent natural killer cell-mediated cytotoxicity engendered by a kinase-inactive HER2 adenovirus-based vaccination mediates resistance to breast tumors. Cancer Res. 70:7431-7441. (VPK-100)

10.Sen, P. et al. (2010). Zinc modulates the interaction of protein C and activated protein C with endothelial protein C receptor. J. Biol. Chem. 285:20410-20420. (VPK-100)

11.Sabbatini, M. E. et al. (2010). CCK activates RhoA and Rac1 differentially through G-alpha-13 and G-alpha-q in mouse pan-creatic acini. Am. J. Physiol. Cell Physiol. 298:C592-C605. (VPK-100)

12.Lam, Y.W. et al. (2010). Proteomics analysis of the nucleolus in adenovirus-infected cells. Mol. Cell. Proteomics 9:117-130. (VPK-100)

Selection Guide for ViraBind™ Adenovirus Purification Kits

ViraBind™ Adenovirus Miniprep Kit

ViraBind™ Adenovirus Purification Kit

Purification Method Spin column Syringe filter

Purification Time 30 minutes 30 minutes

Capacity/Prep (Viral Particles) 1 x 1011 VP 2.5 x 1012 VP

Capacity/Prep (Supernatant) One T75 flask

or one 10cm dish Four T75 flasks



55

QuickTiter™ Adenoviral Titer & Quantitation Kits

QuickTiter™ Adenovirus Titer Immunoassay Kit. 293AD cells (p. 42) were infected with different dilutions of purified Ad-β-Gal for 48 hours. Immunostaining was performed according to the assay protocol. X-gal staining was performed with β-Galactosidase Stain-ing Kit (p. 106).

Accurate measurement of virus titer is critical for viral gene delivery. Traditional plaque-forming unit (PFU) assays are long and have high inter-assay variability. The QuickTiter™ Adenovirus Titer Kits provide a complete system to functionally titer virus infectivity with greater accuracy in a fraction of the time. The assays may be used with any adenovirus system that can amplify in 293 cells. Assays are available for ICC staining or 96-well ELISA. For a quick test of physical titer, our QuickTiter™ Adenovirus Quantitation Kit measures the concentra-tion of your adenovirus prep in about one hour.

Faster, More Accurate and Precise: Compared to traditional plaque-forming unit assays

User-Friendly Protocol: No agar overlay steps Versatile: Recognize all 41 adenovirus serotypes


QuickTiter™ Adenovirus Titer Immunoassay Kit ICC Staining 100 Assays VPK-109

QuickTiter™ Adenovirus Titer ELISA Kit Colorimetric 2 x 96 Assays VPK-110

QuickTiter™ Adenovirus Quantitation Kit Fluorometric 20 Assays VPK-106

Recent Product Citations 1. Smith, M. et al. (2010). PRDM1/Blimp-1 controls effector cyto-

kine production in human NK cells. J. Immunol. 185:6058-6067. (VPK-106)

2. Reiter, C.E.N. et al. (2010). Green tea polyphenol epigallocate-chin gallate reduces endothelin-1 expression and secretion in vascular endothelial cells: roles for AMP-activated protein kinase, Akt, and FOXO1. Endocrinology 151:103-114. (VPK-106)

3. Wilkins, H. et al. (2013). Mitochondrial glutathione transport is a key determinant of neuronal susceptibility to oxidative and nitrosative stress. J. Biol. Chem. 288:5091-5101. (VPK-109)

4. Scallan, C. et al. (2013). An adenovirus-based vaccine with a double-stranded RNA adjuvant protects mice and ferrets against H5N1 avian influenza in oral delivery models. Clin. Vaccine Immunol. 20:85-94. (VPK-109)

5. Xiong, X. et al. (2012). The autophagy-related gene 14 (Atg14) is regulated by forkhead box O transcription factors and cir-cadian rhythms and plays a critical role in hepatic autophagy and lipid metabolism. J. Biol. Chem. 287:39107-39114. (VPK-109)

6. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phos-phorylation of vascular endothelial cadherin induced by inva-sive breast cancer cells. J. Biol. Chem. 287:32981-32992. (VPK-109)

7. Hisamitsu, T. et al. (2012). Na+/H+ exchanger 1 directly binds to calcineurin A and activates downstream NFAT signaling, leading to cardiomyocyte hypertrophy. Mol. Cell Biol. 32:3265-3280. (VPK-109)

8. Lee, S. et al. (2012). Adiponectin abates diabetes-induced endothelial dysfunction by suppressing oxidative stress, adhe-sion molecules, and inflammation in type 2 diabetic mice. Am. J. Heart Circ. Physiol. 303:H106-H115. (VPK-109)

QuickTiter™ Adenovirus Titer Immunoassay Kit

QuickTiter™ Adenovirus Titer ELISA Kit

QuickTiter™ Adenovirus Quantitation Kit

Functional or Physical Titer

Functional (Infectious units)

Functional (Infectious units)

Physical (Viral particles)

Assay Time 2.5 days 2.5 days 45-60 minutes

Assay Principle Antibody-based Antibody-based Total nucleic acid content

Detection Method Immunocytochemical

staining Colorimetric (ELISA)

plate reader Fluorescence plate reader

Key Benefit Accuracy Accuracy Speed

Selection Guide for QuickTiter™ Adenoviral Quantitation Kits



56

Rapid Replication Competent Adenovirus (RCA) Assay Kit


Rapid RCA Assay Kit 30 Assays VPK-111

5 x 30 Assays VPK-111-5 ICC Staining

This kit uses the assay principles of the QuickTiter™ Adenovirus Titer Immunoassay Kit (see page 47), but is designed specifically to measure the level of repli-cation-competent virus in your adenoviral prep.

Adenovirus infection of target cells is mediated largely by the coxsackievirus-adenovirus receptor (CAR). Generally adenoviral transduction of many immortalized cell lines proceeds with a high level of efficiency. However, in many primary cells this re-ceptor is either absent or present at extremely low-levels. This can reduce the efficiency of adenovirus transduction into your cell of choice. ViraDuctin™ Adenovirus Transduction Reagent is designed specifically to increase the efficiency of adenoviral transduction, without regard to the level of CAR expression on the surface of the target cells.

Higher Transduction Efficiency: Up to 12-fold increase in adenoviral uptake

User-Friendly: Short incubation step prior to host cell infection

Versatile: Ideal for target cells expressing little or no CAR, but may also improve transduction efficiency for CAR-expressing cells


10 Transductions AD-200

50 Transductions AD-201 ViraDuctin™ Adenovirus Transduction Reagent (CAR-Independent)

Enhanced Transduction using ViraDuctin™ Adenovirus Trans-duction Reagent. Infection of NIH3T3 cells with recombinant Ad-ß-gal (ADV-002). Top: X-gal staining under microscope. Bottom: scoring of infection with ViraDuctin™ reagent as a percentage of infection with control.

0

200

400

600

800

1000

1200

1400

Control ViraDuctin™

X-G

al S

tain

ing

Po

sit

ive

(%

)

ViraDuctin™ Adenovirus Transduction Reagent, CAR-Independent

Recent Product Citations 1. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phos-

phorylation of vascular endothelial cadherin induced by inva-sive breast cancer cells. J. Biol. Chem. 287:32981-32992.

2. Ackerman, W. et al (2008). Nuclear Factor-kappa B regulates inducible prostaglandin E synthase expression in human am-nion mesenchymal cells. Biol. Reprod. 78:68-76.

3. Monick, M. et al. (2008). Constitutive ERK MAPK activity regulates macrophage ATP production and mitochondrial integrity. J. Immunol. 180:7485-7496.

*Based on using 6-well plates or 35mm culture dishes; may also be used with 96-,24- or 12-well plates or 60mm or 100mm dishes.

Faster Results: 2.5 days vs. 10 days with plaque assay

Versatile: Recognizes all 41 adenovirus serotypes

Immunostaining of Wild Type Ad5 using the Rapid RCA Assay Kit.


57

VIRAL EXPRESSION Lentiviral Expression

Lentiviral Expression Kits & Reagents

As a sub-class of retroviruses, lentiviruses based on HIV-1 have the unique advantage of being able to infect both proliferating and non-proliferating cells, and they can be used for both transient and stable gene expression. We offer a complete workflow solution to your lentiviral expression studies:

Concentration / Purification Kits Quantitation / Titer Kits Transduction Reagents

Expression Systems & Vectors Premade Controls Viral Host Cell Line

ViraSafe™ Lentiviral Expression Systems

Lentiviruses based on HIV-1 may infect both dividing and non-dividing cells. Recently developed third-generation lentiviral expression systems have re-duced the risk of creating replication-competent virus upon recombination, but the risk is still present. Our ViraSafe™ Lentiviral Expression Systems pro-vide a safer and more flexible method to package your lentivirus, even compared to other third-generation lentivirus systems.

Safer: 80-90% less sequence homology compared to other 3rd-generation lentiviral systems; ecotropic systems provide even more safety*

High Titers: Incorporates elements that provide titers comparable to other 3rd-generation systems

Flexible: Packaging vectors provided separately for increased safety and optimization of vector ratios

Lentivirus Production using the ViraSafe™ Lentiviral Expression System.

*Lentiviruses made with a ViraSafe™ Ecotropic Expression System will only readily infect mouse and rat cells. Pan-tropic lentiviruses are VSVG-pseudotyped and may infect cells of any species.

ViraSafe™ Lentiviral Technology is available in three formats (see next two pages for ordering information): Complete Expression Systems: Include 3 packag-

ing plasmids, expression vector and control vector Packaging Systems: Include the 3 individual pack-

aging plasmids; ideal if you already have a 3rd-generation lentiviral expression construct

Expression Vectors: 11 cloning vectors to choose from; compatible with any 2nd or 3rd generation packaging system, but produce the highest titers with the ViraSafe™ packaging system


58


Product Name Envelope Size Catalog Number

ViraSafe™ Universal Lentiviral Expression System (Promoterless) Ecotropic 1 Kit VPK-211-ECO

Pantropic (VSVG) 1 Kit VPK-211-PAN

ViraSafe™ Lentiviral Expression System (Puro) Ecotropic 1 Kit VPK-212-ECO


ViraSafe™ Lentiviral Expression System (Neo) Ecotropic 1 Kit VPK-213-ECO


ViraSafe™ Lentiviral Expression System (Hygro) Ecotropic 1 Kit VPK-214-ECO


ViraSafe™ Lentiviral Bicistronic Expression System (Puro) Ecotropic 1 Kit VPK-215-ECO


ViraSafe™ Lentiviral Bicistronic Expression System (Neo) Ecotropic 1 Kit VPK-216-ECO


ViraSafe™ Lentiviral Bicistronic Expression System (Hygro) Ecotropic 1 Kit VPK-217-ECO


ViraSafe™ Lentiviral Bicistronic Expression System (GFP) Ecotropic 1 Kit VPK-218-ECO


ViraSafe™ Lentiviral Bicistronic Expression System (Blasticidin) Ecotropic 1 Kit VPK-219-ECO


Ecotropic 1 Kit VPK-221-ECO


ViraSafe™ shRNA Lentiviral Expression System (GFP) Ecotropic 1 Kit VPK-222-ECO


ViraSafe™ shRNA Lentiviral Expression System (Puro)

ViraSafe™ Lentiviral Expression Systems, continued

Complete ViraSafe™ Expression Systems include three individual packaging plasmids, an expression vector, and a control vector. Choose an ecotropic system for infection of mouse or rat cells, or a pan-tropic system to produce VSVG-pseudotyped lenti-virus for infection of cells from any species.

ViraSafe™ Lentiviral Packaging Systems

ViraSafe™ Packaging Systems contain 3 packaging plasmids for use with any 3rd-generation lentiviral expression vector. These systems are perfect if you already have a lentiviral construct containing your gene of interest.

Product Name Envelope Size Catalog Number

ViraSafe™ Lentiviral Packaging System Ecotropic 1 Kit VPK-205

1 Kit VPK-206 Pantropic (VSVG)


Recent Product Citation Davis, M. et al. (2013). RAC1P29S is a spontaneously activating cancer-associated GTPase. PNAS 110:912-917. (VPK-214-PAN)

Recent Product Citation Vogt, J. et al. (2014). Protein associated with SMAD1 (PAWS1/FAM83G) is a substrate for type I bone morphogenetic protein receptors and modulates bone morphogenetic protein signaling. Open Bio. 4:130210. (VPK-206)

Recent Product Citations 1. Rossello, R.A. et al. (2013). Mammalian genes induce partially reprogrammed pluripotent stem cells in non-mammalian vertebrate and

invertebrate species. eLife Sci. 2:e00036. 2. Dillahunt, S. et al. (2013). Usage of sphingosine kinase isoforms in mast cells is species and/or cell type determined. J. Immunol.

190:2058-2067.

59


293LTV Lentiviral Cell Line


293LTV Cell Line 1 x 106 Cells LTV-100

ViraSafe™ Lentiviral Expression Vectors


pSMPUW Universal Lentiviral Expression Vector (Promoterless) 10 µg VPK-211

pSMPUW-Puro Lentiviral Expression Vector 10 µg VPK-212

pSMPUW-Neo Lentiviral Expression Vector 10 µg VPK-213

pSMPUW-Hygro Lentiviral Expression Vector 10 µg VPK-214

pSMPUW-IRES-Puro Lentiviral Expression Vector 10 µg VPK-215

pSMPUW-IRES-Neo Lentiviral Expression Vector 10 µg VPK-216

pSMPUW-IRES-GFP Lentiviral Expression Vector 10 µg VPK-218

pSMPUW-IRES-Bsd Lentiviral Expression Vector 10 µg VPK-219

pSMPUW-U6-Puro Lentiviral Expression Vector 10 µg VPK-221

pSMPUW-IRES-Hygro Lentiviral Expression Vector 10 µg VPK-217

pSMPUW-U6-GFP Lentiviral Expression Vector 10 µg VPK-222

Cloning Capacity

9.4 kb

7.9 kb

7.7 kb

7.4 kb

7.8 kb

7.5 kb

7.3 kb

7.6 kb

7.9 kb

7.7 kb

7.6 kb

These lentiviral expression vectors may be used with any 2nd or 3rd generation lentiviral packaging sys-tem, but best results are achieved when used with our ViraSafe™ Lentiviral Packaging Systems.

Lentiviral Control Plasmids


pLenti-GFP Lentiviral Control Vector 10 µg LTV-400

pSMPUW-GFP-Puro Lentiviral Control Vector 10 µg LTV-401

pSMPUW-MNDnLacZ Lentiviral Control Vector 10 µg LTV-402

pLenti-RFP-Puro Lentiviral Control Vector 100 µL LTV-403

Premade Reporter Lentivirus Controls

Product Name Concentration Size Catalog Number

GFP Lentivirus Control 1 x 106 TU/mL 200 µL LTV-300

RFP Lentivirus Control 1 x 106 TU/mL 200 µL LTV-301


Recent Product Citations 1. Sankaran, V.G. et al. (2011). MicroRNA-15a and -16-1 act via

MYB to elevate hemoglobin expression in human trisomy 13. PNAS 108(4):1519-1524. (VPK-212)

2. Davis, M. et al. (2013). RAC1P29S is a spontaneously activating cancer-associated GTPase. PNAS 110:912-917. (VPK-214)

Our 293LTV cell line was selected from the parental 293T cell line for firmer attachment to culture plates and larger, rounder morphology for greater lentiviral production.

Assay Principle for the QuickTiter™ Lentivirus Titer Kit. Lenti-virus particles are packaged with p24 protein, but additional free p24 protein is present in viral supernatant. A traditional p24 ELISA detects both sources of p24 which overestimates viral titer. The QuickTiter™ Lentivirus Titer Kit uses technology to pull the virus out of solution prior to quantitation for a more accurate viral titer.

60


QuickTiter™ Lentivirus Titer / Quantitation Kits

Measuring lentiviral titer is important prior to infection of your target cells, and one of the most published methods is the p24 ELISA. Our traditional p24 ELISA kit provides a quick, convenient way to quantify the concentration of your HIV-1 based lentivirus. One disadvantage of using a traditional p24 ELISA to quantify lentivirus is the overexpression of p24 during lentiviral packaging. Free p24 protein may account for a substantial portion of total p24 in lentiviral super-natant. The traditional p24 ELISA detects both virus-associated p24 and free p24 generated by 293T cells during transient transfection. Our QuickTiter™ Lenti-virus Titer Kit minimizes the overestimation of p24 in lentivirus supernatant. Our proprietary technology separates the lentivirus-associated p24 from free p24 protein prior to performing the ELISA. If you need a very quick estimate of your lentiviral concentration, try the QuickTiter™ Lentivirus Quanti-tation Kit. This kit specifically measures the viral nu-cleic acid content of purified virus or unpurified viral supernatant. This method is ideal for a quick meas-urement of viral titer, either before or after purification of your lentivirus.

More Accurate: Exclusive technology in Quick-Titer™ Lentivirus Titer Kit minimizes overestima-tion of virus titer

User-Friendly: Read results on a standard microplate reader

Selection Guide for QuickTiter™ Lentivirus Quantitation & Titer Kits

QuickTiter™ Lentivirus Titer Kit

(Lentivirus-Associated p24 ELISA)

QuickTiter™ Lentivirus Quantitation Kits

(Traditional p24 ELISA)

QuickTiter™ Lentivirus

Quantitation Kit

Assay Principle p24 ELISA with proprietary

technology to separate free p24 from viral p24

p24 ELISA Measures

nucleic acid content

Suitable Viruses Recombinant HIV-1 Recombinant or native

HIV-1 HIV-1, FIV, SIV

Detection Method Colorimetric (ELISA)

plate reader Colorimetric (ELISA)

plate reader Fluorescence plate reader

Key Benefit Accuracy Most Published Speed (45-60 min.)


QuickTiter™ Lentivirus Titer / Quantitation Kits, continued

0

0.2

0.4

0.6

0.8

1

1.2

Culture medium GFP LentiviralSupernatant

OD

45

0 n

m

p24 Titer of GFP Lentiviral Supernatant. GFP lentiviral construct was cotransfected with a packaging mix into 293 cells. The condi-tioned medium was harvested 48 hrs after transfection and used to further infect 293 cells. The p24 level of the diluted lentiviral super-natant (1:10 dilution) was determined as described in the assay protocol.

61


0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

0 25 50 75 100

p24 (ng/mL)

OD

45

0n

m

p24 in supernatant

p24 in pellet


QuickTiter™ Lentivirus Titer Kit (Lentivirus-Associated HIV p24 ELISA) Colorimetric 96 Assays VPK-107

5 x 96 Assays VPK-107-5

QuickTiter™ Lentivirus Quantitation Kit (HIV-1 p24 ELISA) Colorimetric 96 Assays VPK-108-H

5 x 96 Assays VPK-108-H-5

QuickTiter™ Lentivirus Quantitation Kit Fluorometric 20 Assays VPK-112

Free p24 Does not Complex with ViraBind™ Reagents. Recom-binant p24 was diluted in culture medium and treated with Vira-Bind™ Lentivirus Reagents A and B found in the QuickTiter™ Lentivirus Titer Kit. The amount of p24 in the supernatant and the pellet was measured according to the assay protocol.

Recent Product Citations 1. Belaner, K. et al. (2013). Binding of RNA by APOBEC3G con-

trols deamination-independent restriction of retroviruses. J. Exp. Biol. 216:2213-2220. (VPK-107)

2. Yu, X. et al. (2012). Identification of Hepatitis B virus inhibitors targeting different aspects of infection using a cell-based assay. Antimicrob. Agents Chemother. 56:6109-6120. (VPK-107)

3. Walker, K. et al. (2012). Depletion of GGA1 and GGA3 mediates postinjury elevation of BACE1. J. Neurosci. 32:10423-10437. (VPK-107)

4. Zhou, B. et al. (2012). Interactions between ß-catenin and Transforming growth factor-ß signaling pathways mediate epithelial-mesenchymal transition and are dependent on the transcriptional co-activator cAMP-response element-binding protein (CREB)-binding. J. Biol. Chem. 287:7026-7038. (VPK-107)

5. Nedelec, A.D. et al. (2012). Noonan Syndrome-causing SHP2 mutants inhibit insulin-like growth factor 1 release via growth hormone-induced ERK hyperactivation, which contributes to short stature. PNAS 109:4257-4262. (VPK-107)

6. Lavender, H. et al. (2012). In vitro characterization of the activity of PF-05095808, a novel biological agent for Hepatitis C virus therapy. Antimicrob. Agents Chemother. 56:1364-1375. (VPK-107)

7. Keck, Z.Y. et al. (2011). Mapping a region of Hepatitis C virus E2 that is responsible for escape from neutralizing antibodies and a core CD81-binding region that does not tolerate neutrali-zation escape mutations. J. Virol. 85:10451-10463. (VPK-107)

8. Sanchez-Antequera, Y. et al. (2011). Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells. Blood 117:e171-e181. (VPK-107)

9. Yi, S.H. et al. (2014). Foxa2 acts as a co-activator potentiating expression of the Nurr1-induced DA phenotype via epigenetic regulation. Development 141:761-772. (VPK-108-H)

10. Smith, B. et al. (2013). Targeting the PyMT oncogene to diverse mammary cell populations enhances tumor heterogeneity and generates rare breast cancer subtypes. Genes & Cancer 10.1177/1947601913475359. (VPK-108-H)

11. Iftikhar, M. et al. (2011). Lysyl oxidase-like-2 (LOXL2) is a major isoform in chondrocytes and is critically required for differentia-tion. J. Biol. Chem. 286:909-918. (VPK-108-H)

12. Agrawal-Gamse, C. et al. (2010). Yeast-elicited cross-reactive to HIV Env glycans efficiently neutralize virions expressing exclu-sively high mannose N-linked glycans. J. Virol. 85(1):470-480. (VPK-108-H)

13. Rossello, R.A. et al. (2013). Mammalina genes induce partially reprogrammed pluripotent stem cells in non-mammalian verte-brate and invertebrate species. eLife Sci. 2:e00036. (VPK-112)

14. Fan, X. et al. (2012). Transient, inducible, placenta-specific gene expression in mice. Endocrinology 153:5637-5644. (VPK-112)

15. Veeraraghavalu, K. et al. (2010). Presinilin 1 mutants impair the self-renewal and differentiation of adult murine subventricular zone-neuronal progenitors via cell-autonomous mechanisms involving notch signaling. J. Neurosci. 30:6903-6915. (VPK-112)



62

ViraBind™ Lentivirus Concentration & Purification Kits

Lentivirus Concentration and Purification Procedure.

Ultracentrifugation methods used for lentiviral super-natants are tedious and time-consuming and usually only partially purify your virus. Alternatively, filter-based methods have low sample capacity and do not substantially concentrate the virus. ViraBind™ Lentivirus Concentration and Purification Kits produce purified lentivirus with extremely high titer without the need for ultracentrifugation. The virus is pelleted from solution using our proprietary isola-tion technology, then resuspended in a smaller vol-ume. The resuspended lentivirus is then applied to either a purification column or a dialysis device for purification. If desired, the purified virus may be fur-ther concentrated using a centrifugal concentrator.

Fast: Obtain purified virus in about 4-6 hours with column-based kits and 10-24 hours with dialysis-based kits

High Titer: Concentrate up to 500-fold to as high as 108-1010 TU/ml, sufficient for in vivo studies

High Yield: Recover >60%


ViraBind™ Lentivirus Concentration and Purification Kit (100 ml/prep)

2 Preps VPK-090

5 Preps VPK-091

25 Preps VPK-091-5

ViraBind™ PLUS Lentivirus Concentration and Purification Kit (50 ml/prep) 2 Preps VPK-095

2 Preps VPK-096

10 Preps VPK-096-5 ViraBind™ PLUS Lentivirus Concentration and Purification Mega Kit (500 ml/prep)

Selection Guide for Lentivirus Concentration & Purification Kits

ViraBind™ Lentivirus

Concentration and Purification Kit

ViraBind™ PLUS Lentivirus


ViraBind™ PLUS Lentivirus

Concentration and Purification Mega Kit

Purification Method

Proprietary Reagent Cocktail + Purification Column

Proprietary Reagent Cocktail + Dialysis


Total Time 6-8 hours 10-24 hours 10-24 hours

Capacity per Prep(Supernatant)

100 mL 50 mL 500 mL



63

ViraDuctin™ Lentivirus Transduction Kit


40 Transductions LTV-200

200 Transductions LTV-201 ViraDuctin™ Lentivirus Transduction Kit

Transduction of 293AD and HT-1080 Cells. 293AD cells (p. 36) and HT-1080 cells were each seeded at 50,000 cells/well in a 24-well plate overnight. Cells were infected with GFP lentivirus for 48 hours in the presence of no additive (left), Polybrene® (middle) or the ViraDuctin™ reagent cocktail (right).

Transduction Efficiencies in Various Cell Lines. NIH3T3 cells, HeLa cells, our own 293AD cells (page 36) and HT-1080 cells were each seeded at 50,000 cells/well in a 24-well plate overnight. Cells were infected with GFP lentivirus for 48 hours in the presence of Polybrene® or ViraDuctin™ Lentivirus Transduc-tion Kit. For each cell line, fluorescence levels using the Vira-Ductin™ system are depicted relative to a normalized fluores-cence of 100 for Polybrene®.

Lentivirus transduction efficiency is typically low. Ad-ditives such as Polybrene® can boost transduction efficiencies, but even then only a small fraction of len-tiviral vectors can transduce many target cell lines. Our ViraDuctin™ Lentivirus Transduction Kit provides superior transduction efficiencies in a variety of cell lines, even when compared to transductions in the presence of Polybrene®.

Higher Transduction Efficiency: 2-6x higher in many cell lines compared to Polybrene

More Robust: Useful for transduction of nonper-missive cells, including primary cells and stem cells

Polybrene is a registered trademark of Abbott Laboratories.

*Based on a 24-well plate. Can also be used with 96-well, 12-well or 6-well plates, as well as 60mm or 100mm dishes. See product insert.

0

100

200

300

400

500

600

700

NIH3T

3

HeLa

293

HT-108

0No

rmal

ized

GF

P F

luo

resc

ence

Polybrene

ViraDuctin™


Recent Product Citations 1. Rossello, R.A. et al. (2013). Mammalina genes induce partially

reprogrammed pluripotent stem cells in non-mammalian verte-brate and invertebrate species. eLife Sci. 2:e00036.

2. McEachron, T.A. et al. (2010). Protease-activated receptors mediate crosstalk between coagulation and fibrinolysis. Blood 116:5037-5044.

3. Zemskova, M. et al. (2010). p53-dependent induction of prostate cancer cell senescence by the PIM1 protein kinase. Mol. Cancer Res. 8:1126-1141.

VIRAL EXPRESSION Retroviral Expression

64

Retroviral Expression Kits & Reagents

Traditional retroviral vectors based on MMLV are useful for integrating genetic material into the host cell genome. However, retrovirus titer tends to be significantly lower than that of adenovirus, which can lead to a lower infection efficiency. Our retroviral reagents and kits incorporate technologies that increase your chances of successful retroviral expression. We offer a comprehensive solution from start to finish:

Gene-Specific Retroviral Vectors Concentration / Purification Kits Quantitation / Titer Kits Transduction Reagents

Retroviral Expression Systems Retroviral Packaging Cell Lines Retroviral Cloning & Expression

Vectors



Platinum-E Retroviral Packaging Cell Line, Ecotropic >3 x 106 cells RV-101

Platinum-A Retroviral Packaging Cell Line, Amphotropic >3 x 106 cells RV-102

Platinum-GP Retroviral Packaging Cell Line, Pantropic >3 x 106 cells RV-103

pVSV-G Packaging Vector 10 µg RV-110

Platinum Retroviral Packaging Cell Lines

Recent Product Citations 1. Schmidt, T. et al. (2013). CXCR4 promotes B cell egress from

Peyer’s patches. J. Exp. Med. 10.1084/jem.20122574. (RV-101) 2. Wahlestedt, M. et al. (2013). An epigenetic component of hemato-

poietic stem cell aging amenable to reprogramming into a young state. Blood 121:4257-4264. (RV-101)

3. Zhong, S. et al. (2013). T-cell receptor affinity and avidity defines antitumor response and autoimmunity in T-cell immunotherapy. PNAS 110:6973-6978. (RV-101)

4. Nam, Y.J. et al. (2013). Reprogramming of human fibroblasts to-ward a cardiac fate. PNAS 110:5588-5593. (RV-102)

5. Hrdlickova, R. et al. (2012). Alternatively spliced telomerase re-verse transcriptase variants lacking telomerase activity stimulate cell proliferation. Mol. Cell Biol. 32:4283-4296. (RV-102)

6. Nowakowski, T. et al. (2013). MicroRNA-92b regulates the devel-opment of intermediate cortical progenitors in embryonic mouse brain. PNAS 110:7056-7061. (RV-103)

7. Cavnar, P.J. et al. (2012). The actin regulatory protein HS1 inter-acts with Arp2/3 and mediates efficient neutrophil chemotaxis. J. Biol. Chem. 287:25466-25477. (RV-103)

Generate high titers of recombinant retrovirus with a single plasmid transfection* using these extremely powerful, stable cell lines. Platinum Retroviral Packag-ing Cells are based on the 293T cell line and exhibit greater stability and produce higher yields of retroviral structure proteins, resulting in higher retroviral titers. The Platinum cell lines were invented in the laboratory of Dr. Toshio Kitamura at the University of Tokyo and are available exclusively from Cell Biolabs. They were first described in the following paper: Morita, S. et al. (2000). Gene Therapy 7:1063-1066.

Plat-A Cells(Amphotropic)

Plat-E Cells(Ecotropic)

Plat-GP Cells(Pantropic*)

Human +++ N.S. +++

Mouse +++ +++ +++

Rat +++ +++ +++

Monkey +++ N.S. +++

Cat +++ N.S. +++

Dog +++ N.S. +++

Hamster + N.S. +++

Bird N.S. N.S. +++

Fish N.S. N.S. +++

Frog N.S. N.S. +++

Insect N.S. N.S. +++

Mollusk N.S. N.S. +++

*Plat-GP cells must be co-transfected with a pantropic envelope protein such as VSV-G. N.S. = Not Suitable

Suitability of Platinum Retroviral Packaging Cell Lines by Host Species.

Not sure which Platinum Expression System is right for you? See the table below for a selection guide based on the host species of your target cell.



Platinum Retroviral Packaging Cells and Expression Systems

Our Platinum Retroviral Expression Systems incorpo-rate superior packaging cell lines and vector technolo-gies to produce high-titer virus with a single plasmid transfection. Each Platinum Expression System in-cludes one of our exclusive Platinum Packaging Cell Lines which stably express the gag and pol genes. In the Ecotropic and Amphotropic systems, the packag-ing cells also express the envelope protein.* Simply clone your gene of interest into the vector provided and transfect into the Platinum cells. Platinum Retroviral Expression Systems contain eve-rything you need to generate your recombinant retro-virus: packaging cell line, expression vector, and GFP control vector. Our pantropic systems also contain a VSVG envelope vector.

Retrovirus Production Using the Platinum Expression Systems (Ecotropic and Amphotropic).

Higher Viral Yields: Average titer 107 infectious units/mL with transient transfection

Longer Stability: Expression up to 4 months in the presence of drug selection

Optimized Systems: 3 packaging cell lines for in-fection of various species; 3 vector backbones (two specifically for infection of stem cells)

Flexible: Order complete systems or cells and vec-tors separately

Product Name Expression Vector Packaging Cell Catalog Number

Platinum Retroviral Expression System, Ecotropic pMXs-Puro Plat-E VPK-300

Platinum Retroviral Expression System, Amphotropic pMXs-Puro Plat-A VPK-301

Platinum Retroviral Expression System, Pantropic pMXs-Puro Plat-GP VPK-302

Platinum ES/EC Retroviral Expression System, Ecotropic pMCs-Puro Plat-E VPK-303

Platinum ES/EC Retroviral Expression System, Amphotropic pMCs-Puro Plat-A VPK-304

Platinum ES/EC Retroviral Expression System, Pantropic pMCs-Puro Plat-GP VPK-305

Platinum HSC Retroviral Expression System, Ecotropic pMYs-Puro Plat-E VPK-306

Platinum HSC Retroviral Expression System, Amphotropic pMYs-Puro Plat-A VPK-307

Platinum HSC Retroviral Expression System, Pantropic pMYs-Puro Plat-GP VPK-308

Recent Product Citations 1. Aoi, N. et al. (2012). 1a,25-dihydroxyvitamin D3 modulates the hair

-inductive capacity of dermal papilla cells: therapeutic potential for hair regeneration. Stem Cells Trans Med. 1:615-626. (VPK-301)

2. Wang, N. et al. (2013). Lacritin rescues stressed epithelia via rapid forkhead box O3 (FOXO3)-associated autophagy that restores metabolism. J. Biol. Chem. 288:18146-18161. (VPK-302)

3. Tanaka, T. et al. (2012). Anthracycline inhibits recruitment of hy-poxia-inducible transcription factors and suppresses tumor cell migration and cardiac angiogenic response in the host. J. Biol. Chem. 287:34866-34882. (VPK-302)

*Pantropic systems require co-transfection with the provided VSVG envelope vector.

66



Retroviral Cloning & Expression Vectors

Our Retroviral Expression Vectors are based on backbones derived from Moloney murine leukemia virus (MMLV). We offer the traditional pBABE system and the novel pMXs system, which has been shown to be useful in induced pluripotent stem cell (iPS) studies. pMYs vectors are optimal for use with hema-topoietic stem cells, and pMCs vectors are optimal for ES and EC cells. All cloning vectors are supplied as 10 µg in TE buffer.

Vector Name Cloning Capacity Catalog Number

pMXs-U6-GFP 5 kb RTV-071

pMXs-U6-Puro 5.1 kb RTV-070

pMXs-U6-Puro-shGFP RTV-055

pMXs-U6-Puro-shLuc RTV-056

Recent Product Citations 1. Duran, P.P. et al. (2012). UNG shapes the specificity of AID-

induced somatic hypermutation. J. Exp. Med. 209:1379-1389. (RTV-001-HYGRO)

2. Miyoshi, N. et al. (2010). Defined factors induce reprogramming of gastrointestinal cancer cells. PNAS 107:40-45. (RTV-010)

3. Parikh, C. et al. (2012). Disruption of PH-kinase domain interac-tions leads to oncogenic activation of AKT in human cancers. PNAS 109:19368-19373. (RTV-012)

4. Zhang, Q. et al. (2013). TNF-a impairs differentiation and func-tion of TGF-ß-induced Treg cells in autoimmune diseases through Akr and Smad3 signaling pathway. J. Mol. Cell Biol. 10.1093/jmcb/jms063. (RTV-013)

5. Sugatani, T. et al. (2011). A microRNA expression signature of osteoclastogenesis. Blood 117:3648-3657. (RTV-014, RTV-016)

Retroviral Cloning Vectors for General Gene Expression (driven by 5’ LTR)


pBABEhygro 5.6 kb RTV-001-HYGRO

pBABEneo 5.9 kb RTV-003

pBABEpuro 6 kb RTV-001-PURO

pBABEzeo 6.3 kb RTV-004

pMXs 5.4 kb RTV-010

pMXs-IRES-Bsd 5.6 kb RTV-016

pMXs-IRES-GFP 5.3 kb RTV-013

pMXs-IRES-Neo 5.2 kb RTV-015

pMXs-IRES-Puro 5.4 kb RTV-014

pMXs-Neo 3.8 kb RTV-011

pMXs-Puro 4.4 kb RTV-012

pMZs 5.3 kb RTV-030

Retroviral Cloning Vectors with Strong Promoters for Overexpression


pMXs-CAG 5.2 kb RTV-064

pMXs-CMV 5.5 kb RTV-065

pMXs-EF1 5.5 kb RTV-063

pMXs-EF1-Bsd 4.2 kb RTV-062

pMXs-EF1-GFP 3.9 kb RTV-061

pMXs-EF1-Puro 4 kb RTV-060

pMXs-SR 5.4 kb RTV-066

Retroviral Cloning Vectors for use with ES/EC Cells


pMCs-IRES-GFP 5.2 kb RTV-040

pMCs-Puro 4.3 kb RTV-041

Retroviral Cloning Vectors for use with Hematopoietic Cells


pMYs 5.2 kb RTV-020

pMYs-IRES-GFP 5.2 kb RTV-021

pMYs-IRES-Neo 5.2 kb RTV-023

pMYs-IRES-Puro 5.4 kb RTV-022

pMYs-Puro 4.3 kb RTV-024

Retroviral Cloning Vector for miRNA


pMXs-miR-GFP/Puro 4.2 kb RTV-017

Retroviral Cloning Vectors for shRNA

Recent Product Citations 1. Malicet, C. et al. (2011). Distinct properties of human HMGN5

reveal a rapidly evolving but functionally conserved nucleosome binding protein. Mol. Cell Biol. 31:2742-2755. (RTV-040)

2. Mochizunki, Y. et al. (2013). Phosphatidylinositol 3-phosphate myotubularin-related protein 6 (MTMR6) is regulated by small GTPase Rab1b in the early secretory and autophagic pathways. J. Biol. Chem. 288:1009-1021. (RTV-041)

Recent Product Citation Mansour, M. et al. (2013). The TAL1 complex targets the FBXW7 tumor suppressor by activating miR-223 in human T cell acute lymphoblastic leukemia. J. Exp. Med. 210:1545-1557. (RTV-017)


67 www.cellbiolabs.com

Retroviral Packaging Vectors and Cells

These constructs are ideal for researchers who prefer a traditional multi-plasmid transfection of 293 cells for packaging recombinant retrovirus.


pCMV-10A1 Envelope Vector 100 µL RV-114

pCMV-Ampho Envelope Vector 100 µL RV-113

pCMV-Eco Envelope Vector 100 µL RV-112

pCMV-Gag-Pol Retroviral Vector 10 µg RV-111

pCMV-VSV-G Envelope Vector 10 µg RV-110

Recent Product Citations 1. Amagai, Y. et al. (2013). Stem cell factor contributes to tumorigenesis of mast cells via an autocrine/paracrine mechanism. J. Leukoc.

Biol. 93:245-250. (RV-110) 2. Okamoto, K. et al. (2012). Dengue virus strain DEN2 16681 utilizes a specific glycochain of syndecan-2 proteoglycan as a receptor. J.

Gen. Virol. 93:761-770. (RV-110, RV-111)


c-Abl pBABEpuro RTV-402

c-Abl-TM pBABEpuro RTV-403

c-Abl (1-565) pBABEpuro RTV-404

c-Abl (1-958) pBABEpuro RTV-405

pBABEhygro RTV-007

pBABEneo RTV-005

pBABEpuro RTV-006

p53 pBABEpuro RTV-401

hTERT

Cell Cycle

Recent Product Citation Huang, J. et al. (2009). Regulation of the leucocyte chemoattrac-tant receptor FPR in glioblastoma cells by cell differentiation. Carcinogenesis 30(2):348-355. (RTV-401)

Reporter Genes


GFP RTV-002

GFP RTV-051

Vector Backbone

pBABE

pMCs

GFP pMX RTV-050

GFP pMYs RTV-052

GFP-Puro pMX RTV-053

Recent Product Citations 1. Hrdlickova, R. et al. (2012). Alternatively spliced telomerase

reverse transcriptase variants lacking telomerase activity stimu-late cell proliferation. Mol. Cell Biol. 32:4283-4296. (RTV-002)

2. Wahlestedt, M. et al. (2013). An epigenetic component of hema-topoietic stem cell aging amenable to reprogramming into a young state. Blood 121:4257-4264. (RTV-050)

Autophagy


GFP-LC3 pMXs RTV-801

This vector is supplied with a separate pMXs-GFP control vector at no additional cost.

These constructs are based on backbones derived from MMLV, Vectors with GFP or stem cell factors are supplied as 10 µg of plasmid in TE buffer. All other vec-tors are supplied as 100 µL of bacterial glycerol stock. Product listing continues on the following pages.

Gene-Specific Recombinant Retroviral Vectors

[email protected]

293RTV Cell Line


293RTV Cell Line >1 x 106 cells RV-100

Our 293RTV cells are derived from the 293 parental cell line, but are selected for firmer attachment to culture plates, faster growth and higher yields of retrovirus produced.

Vector Name Vector Backbone Mutation State Catalog Number

ERK2 pBABEhygro Dominant Negative RTV-109

JNK1 pBABEpuro Dominant Negative RTV-110

MAPKAPK2 pBABEpuro Constitutively Active RTV-118

pBABEpuro Dominant Negative RTV-119

MAPKAPK3 pBABEpuro Constitutively Active RTV-120


MEK1 pBABEhygro Constitutively Active RTV-112

pBABEhygro Dominant Negative RTV-111

MKK3 pBABEpuro Constitutively Active RTV-114


MKK6 pBABEpuro Constitutively Active RTV-116


myr-Akt1 pWZLneo Constitutively Active RTV-125

p38 pBABEhygro Dominant Negative RTV-105




PI3K p110-CAAX pWZLneo Constitutively Active RTV-124

pBABEpuro Constitutively Active RTV-122


Raf1-CAAX pWZLneo Constitutively Active RTV-113

PRAK

MAP Kinase Signaling

68



AUF1 pBABEpuro RTV-305

hnRNPA0 pBABEpuro RTV-310

hnRNP-A2 pBABEpuro RTV-340

HuB pBABEpuro RTV-302

HuC pBABEpuro RTV-303

HuD pBABEpuro RTV-301

HuR pBABEpuro RTV-304

PABP pBABEpuro RTV-307

Stat5A pMXs RTV-330

Stat5A(1*6) pMXs RTV-331

Transcription Regulation

Gene-Specific Recombinant Retroviral Vectors, continued


Stat5A-IRES-GFP pMXs RTV-332

Stat5A(1*6)-IRES-GFP

pMXs RTV-333

Stat5B pMXs RTV-334

Stat5B(1*6) pMXs RTV-335

TIA-1 pBABEpuro RTV-309

TIAR pBABEpuro RTV-308

TTP pBABEpuro RTV-306


Recent Product Citation Yu, Y. et al. (2012). Bcl11a is essential for lymphoid development and negatively regulates p53. J. Exp. Med. 209:2467-2483. (RTV-331)



Gene-Specific Recombinant Retroviral Vectors, continued

Cytoskeleton Regulation

iPS / Stem Cell Factors


4-Vector Set* pMXs RTV-701-C

6-Vector Set** pMXs RTV-709-C

c-Myc pMXs RTV-703

Klf4 pMXs RTV-704

Lin-28 pMXs RTV-710

NANOG pMXs RTV-709


Sox2 pMXs RTV-702


*4-Vector sets contain individual constructs with the following genes: c-Myc, Klf4, Oct-3/4 and Sox2. **6-Vector sets contain individual constructs with the following genes: c-Myc, Klf4, Oct-3/4, Sox2, Lin-28 and NANOG.


4-Vector Set* pMXs RTV-705-C

6-Vector Set** pMXs RTV-711-C

c-Myc pMXs RTV-707

Klf4 pMXs RTV-708

Lin-28 pMXs RTV-712

NANOG pMXs RTV-711


Sox2 pMXs RTV-706


Human iPS Genes Mouse iPS Genes

Proteases and Related Molecules

Recent Product Citation Gutova, M. et al (2008). Urokinase plasminogen activator and urokinase plasminogen activator receptor mediate human stem cell tropism to malignant solid tumors. Stem Cells 26:1406-1413. (RTV-501, RTV-502)


uPA RTV-501

uPAR RTV-502

Vector Backbone

pBABEpuro

pBABEhygro

Recent Product Citation Zhao, B. et al. (2012). TNF-induced osteoclastogenesis and inflammatory bone resorption are inhibited by transcription factor RBP-J. J. Exp. Med. 209:2467-2483. (RTV-101)

Target Name Vector Backbone Mutation State Catalog Number

Cdc42 pBABEhygro L61 RTV-203

K-Ras pBABEpuro N/A RTV-220

pWZLhygro Q61 RTV-221

myr-Rac1 N/A RTV-201

V12 RTV-206

Rac1 pBABEhygro V12 RTV-202

N-Ras pBABEpuro K61 RTV-222

Rac3 pBABEhygro V12 RTV-205

Ras

pBABEpuro V12 RTV-101

pBABEpuro V12C40 RTV-104

pBABEpuro V12G37 RTV-103

pBABEpuro V12S35 RTV-102

RhoA pBABEhygro L63 RTV-204

pBABEpuro


70

ViraBind™ Retrovirus Concentration & Purification Kits

Fast: Obtain purified virus in about 4-6 hours High Titer: Concentrate 500-fold to 109-1010

TU/ml, sufficient for in vivo studies High Yield: Recover >60% High Throughput: Process greater volumes

per prep than filter-based purification methods

Retrovirus Concentration and Purification Procedure.


ViraBind™ Retrovirus Concentration and Purification Kit (100 ml/prep)

2 Preps VPK-130

5 Preps VPK-131

25 Preps VPK-131-5

ViraBind™ PLUS Retrovirus Concentration and Purification Kit (50 ml/prep) 2 Preps VPK-135

2 Preps VPK-136

10 Preps VPK-136-5 ViraBind™ PLUS Retrovirus Concentration and Purification Mega Kit (500 ml/prep)

Selection Guide for Retrovirus Concentration & Purification Kits

ViraBind™ Retrovirus


ViraBind™ PLUS Retrovirus


ViraBind™ PLUS Retrovirus

Concentration and Purification Mega Kit

Purification Method

Proprietary Reagent Cocktail + Purification Column



Total Time 6-8 hours 10-24 hours 10-24 hours

Capacity per Prep(Supernatant)

100 mL 50 mL 500 mL

Ultracentrifugation methods used for lentiviral super-natants are tedious and time-consuming and usually only partially purify your virus. Alternatively, filter-based methods have low sample capacity and do not substantially concentrate the virus. ViraBind™ Retrovirus Concentration and Purification Kits produce purified lentivirus with extremely high titer without the need for ultracentrifugation. The virus is pelleted from solution using our proprietary isola-tion technology, then resuspended in a smaller vol-ume. The resuspended retrovirus is then applied to either a purification column or a dialysis device for purification. If desired, the purified virus may be fur-ther concentrated using a centrifugal concentrator.



71

QuickTiter™ Retrovirus Rapid Quantitation Kit


QuickTiter™ Retrovirus Quantitation Kit Fluorometric 20 Assays VPK-120

Ultra-fast Results: 45-60 minute procedure Convenient: Titer may be measured before purifi-

cation step Sensitive: Limit of detection = 1.5 x 109 VP/mL

from 2 mL of retroviral supernatant

This kit specifically measures the viral nucleic acid con-tent of purified virus or unpurified viral supernatant. This method is ideal for a quick measurement of viral titer, either before or after purification of your retrovirus.

0

100

200

300

400

500

0 400 800 1200

Retroviral RNA (ng)

RF

U (

52

0 n

m)

0

10

20

30

40

50

60

70

80

0 50 100 150

Retroviral RNA (ng)

RF

U (

52

0 n

m)

Retrovirus RNA Standard Curve. The QuickTiter™ Retrovirus RNA Standard was diluted according to the assay protocol. Fluo-rescence was measured on a SpectraMax Gemini XS Fluorometer (Molecular Devices) with a 485 / 538 nm filter set and a 530 nm cutoff.

Assay Procedure for the QuickTiter™ Retrovirus Quantitation Kit.


Recent Product Citation Ito, T. et al. (2012). Stem cell factor programs the mast cell activa-tion phenotype. J. Immunol. 188:5428-5437.


72

ViraDuctin™ Retrovirus Transduction Kit


40 Transductions RV-200

200 Transductions RV-201 ViraDuctin™ Retrovirus Transduction Kit

The efficiency of retrovirus transduction can be low compared to other viruses. The rate at which retrovi-ral vectors bind to cells is controlled mostly by diffu-sion. Additionally, the presence of transduction inhibi-tors such as proteoglycans and glycosaminoglycans in retroviral supernatants can lead to poor gene trans-fer. Additives such as Polybrene® can boost transduc-tion efficiencies, but they do not eliminate these trans-duction inhibitors. Our ViraDuctin™ Retrovirus Transduction Kit pro-vides superior transduction efficiencies even when compared to transductions in the presence of Poly-brene®. A proprietary reagent cocktail forms a super-complex with the retrovirus which is pelleted away from the supernatant, removing detrimental transduc-tion inhibitors that decrease infection efficiency.

More Robust: Removes harmful transduction inhibitors from retroviral supernatant

Higher Transduction Efficiencies: Compared to infections in the presence of Polybrene or no additives

Versatile: Particularly useful for nonpermissive cells including primary cells and stem cells, but may boost transduction rates in a wide variety of cells

Polybrene is a registered trademark of Abbott Laboratories.

*Number of transductions shown is based on use in a 24-well plate. This product may also be used with 96-well, 12-well or 6-well plates, as well as 60mm or 100mm dishes. See product insert for specific details.

Recent Product Citations 1. Gandhi, M. et al. (2012). Homologous chromosomes make con-

tact at the sites of double-strand breaks in genes in somatic G0/G1-phase human cells. PNAS 109:9454-9459.

2. Miyoshi, N. et al. (2010). Defined factors induce reprogramming of gastrointestinal cancer cells. PNAS 107:40-45.


74

MICRORNA ANALYSIS miRNA Clone Collection

miRNASelect™ Precursor Clone Collection (Expression Vectors)

Precursor Name Catalog Number

mmu-let-7a-1 MMU-LET7A-1

hsa-let-7a-2 MIR-LET7A-2

hsa-let-7a-3 MIR-LET7A-3

mmu-let-7b MMU-LET7B

hsa-let7c MIR-LET7C

mmu-let-7c-1 MMU-LET7C-1

mmu-let-7c-2 MMU-LET7C-2

hsa-let-7d MIR-LET7D

mmu-let-7d MMU-LET7D

hsa-let-7e MIR-LET7E

hsa-let-7f-1 MIR-LET7F-1

mmu-let-7f-1 MMU-LET7F-1

mmu-let-7f-2 MMU-LET7F-2

hsa-let-7g MIR-LET7G

mmu-let-7g MMU-LET7G

hsa-let-7i MIR-LET7I

hsa-mir-1-1 MIR-1-1

mmu-mir-1-1 MMU-MIR-1-1

hsa-mir-1-2 MIR-1-2


hsa-mir-7-1 MIR-7-1

hsa-mir-7-2 MIR-7-2

hsa-mir-7-3 MIR-7-3

mmu-mir-7a-1 MMU-MIR-7A-1


mmu-mir-7b MMU-MIR-7B

hsa-mir-9-1 MIR-9-1


hsa-mir-9-2 MIR-9-2

hsa-mir-10a MIR-10A

hsa-mir-10b MIR-10B


hsa-mir-15a MIR-15A

mmu-mir-15a MMU-MIR-15A


miRNASelect™ Expression Vectors contain full miRNA precursor se-quences with constitutive promoter. Human (hsa) vectors contain a

puromycin selection marker Mouse (mmu) vectors contain a

GFP-puromycin fusion


hsa-mir-17 MIR-17

hsa-mir-18a MIR-18A


hsa-mir-18b MIR-18B


hsa-mir-19a MIR-19A


hsa-mir-19b-1 MIR-19B-1

mmu-mir-19b-1 MMU-MIR-19B-1



hsa-mir-20a MIR-20A


hsa-mir-20b MIR-20B


hsa-mir-21 MIR-21

mmu-mir-21 MMU-MIR-21

hsa-mir-22 MIR-22

hsa-mir-23b MIR-23B



hsa-mir-24-2 MIR-24-2



hsa-mir-26a-1 MIR-26A-1




hsa-mir-26b MIR-26B

hsa-mir-27a MIR-27A

hsa-mir-27b MIR-27B





Don’t see your microRNA of interest? Clone your own sequence into one of our

Expression Vectors; see page 80.


Recent Product Citations 1. Dar, A. et al. (2013). The role of miR-18b

in MDM2-p53 pathway signaling and melanoma progression. J. Natl. Cancer Inst. 105:433-442. (MIR-18B)

2. Mallory, M.J. et al. (2011). Signal– and developmental-dependent alternative splicing of LEF1 in T cells is controlled by CELF2. Mol. Cell. Biol. 31:2184-2195. (MIR-23B)

3. Woo, H. et al. (2013). Nucleolin mediates microRNA-directed CSF-1 mRNA deade-nylation but increases translation of CSF-1 mRNA. Mol. Cell Proteomics 12:1661-1667. (MIR-130A, MIR-301A)

4. Starega-Roslan, J. et al. (2010). Stuctural basis of microRNA length variety. Nuc. Acids Res. 10.1093/nar/gkq727. (MIR-148A)

5. Saus, E. et al. (2010). Genetic variants and abnormal processing of pre-miR-182, a circadian clock modulator, in major de-pression patients with late insomnia. Hum. Mol. Genet. 19:4017. (MIR-182)

6. Beezhold, K. et al. (2011). miR-190-mediated downregulation of PHLPP con-tributes to arsenic-induced Akt activation and carcinogenesis. Toxicol. Sci. 123:411-420. (MIR-190)

7. Wang, Y.S. et al. (2012). MicroRNA-195 regulates vascular smooth muscle cell phenotype and prevents neointimal forma-tion. Cardiovasc. Res. 95:5174-526. (MIR-195)

8. Majid, S. et al. (2011). MicroRNA-205 inhibits src-mediated oncogenic pathways in renal cancer. Cancer Res. 71:2611-2621. (MIR-205)

9. Halappanaver, S. et al. (2013). IL-1 recep-tor regulates microRNA-135b expression in a negative feedback mechanism. J. Immunol. 190:3679-3686. (MMU-MIR-135B)

10.Yamamoto, H. et al. (2012). MicroRNA-494 regulates mitochondrial biogenesis in skeletal muscle through mitochondrial transcription factor A and forkhead box j3. Am J. Physiol. Endocrinol. Metab. 303:E1419-1427. (MMU-MIR-494)

75


miRNASelect™ Precursor Clone Collection (Expression Vectors), cont.






hsa-mir-29c MIR-29C

mmu-mir-29c MMU-MIR-29C

hsa-mir-30a MIR-30A


hsa-mir-30b MIR-30B

hsa-mir-30c-1 MIR-30C-1

mmu-mir-30c-1 MMU-MIR-30C-1

hsa-mir-30c-2 MIR-30C-2


hsa-mir-30d MIR-30D

mmu-mir-30d MMU-MIR-30D

hsa-mir-30e MIR-30E

mmu-mir-30e MMU-MIR-30E

hsa-mir-31 MIR-31


hsa-mir-32 MIR-32


hsa-mir-33a MIR-33A

hsa-mir-34c MIR-34C








hsa-mir-95 MIR-95

hsa-mir-96 MIR-96

hsa-mir-28 MIR-28


hsa-mir-29a MIR-29A


hsa-mir-100 MIR-100




hsa-mir-103-1 MIR-103-1


hsa-mir-103-2 MIR-103-2


hsa-mir-105-1 MIR-105-1

hsa-mir-105-2 MIR-105-2

hsa-mir-106a MIR-106A


hsa-mir-106b MIR-106B


hsa-mir-107 MIR-107




hsa-mir-124-2 MIR-124-2








hsa-mir-128-1 MIR-128-1


hsa-mir-128-2 MIR-128-2


hsa-mir-129-1 MIR-129-1



hsa-mir-98 MIR-98






hsa-mir-132 MIR-132












hsa-mir-136 MIR-136


hsa-mir-137 MIR-137


hsa-mir-138-1 MIR-138-1


hsa-mir-138-2 MIR-138-2


hsa-mir-139 MIR-139


hsa-mir-140 MIR-140



hsa-mir-142 MIR-142


hsa-mir-143 MIR-143


hsa-mir-129-2 MIR-129-2






76




hsa-mir-147 MIR-147





hsa-mir-150 MIR-150


hsa-mir-151 MIR-151


hsa-mir-152 MIR-152



hsa-mir-154 MIR-154









hsa-mir-181d MIR-181D




hsa-mir-185 MIR-185


hsa-mir-186 MIR-186

hsa-mir-187 MIR-187



hsa-mir-145 MIR-145








hsa-mir-194-1 MIR-194-1



hsa-mir-195 MIR-195






hsa-mir-197 MIR-197

hsa-mir-198 MIR-198







hsa-mir-200c MIR-200C



hsa-mir-202 MIR-202


hsa-mir-204 MIR-204


hsa-mir-205 MIR-205


hsa-mir-206 MIR-206


hsa-mir-190 MIR-190





hsa-mir-211 MIR-211


hsa-mir-212 MIR-212


hsa-mir-214 MIR-214









hsa-mir-219-1 MIR-219-1



hsa-mir-222 MIR-222



hsa-mir-224 MIR-224










hsa-mir-297 MIR-297







77






hsa-mir-298 MIR-298














hsa-mir-323 MIR-323




hsa-mir-329-1 MIR-329-1


hsa-mir-331 MIR-331


hsa-mir-335 MIR-335




hsa-mir-342 MIR-342





































hsa-mir-433 MIR-433























mmu-mir-466f-1 MMU-MIR-466F-1

mmu-mir-466f-4 MMU-MIR-466F-4

mmu-mir-466g MMU-MIR-466G

mmu-mir-466h MMU-MIR-466H

mmu-mir-466i MMU-MIR-466I

mmu-mir-466j MMU-MIR-466J

mmu-mir-466k MMU-MIR-466K

mmu-mir-466l MMU-MIR-466L




mmu-mir-467f MMU-MIR-467F









78


hsa-mir-603 MIR-603

hsa-mir-605 MIR-605

hsa-mir-606 MIR-606

hsa-mir-608 MIR-608

hsa-mir-609 MIR-609

hsa-mir-610 MIR-610

hsa-mir-613 MIR-613

hsa-mir-616 MIR-616

hsa-mir-619 MIR-619

hsa-mir-620 MIR-620

hsa-mir-628 MIR-628

hsa-mir-630 MIR-630

hsa-mir-633 MIR-633

hsa-mir-635 MIR-635

hsa-mir-636 MIR-636

hsa-mir-637 MIR-637

hsa-mir-641 MIR-641

hsa-mir-643 MIR-643

hsa-mir-645 MIR-645

hsa-mir-649 MIR-649

hsa-mir-651 MIR-651

hsa-mir-652 MIR-652


hsa-mir-653 MIR-653


hsa-mir-654 MIR-654

hsa-mir-658 MIR-658

hsa-mir-659 MIR-659

hsa-mir-665 MIR-665




hsa-mir-600 MIR-600

hsa-mir-601 MIR-601

hsa-mir-602 MIR-602


hsa-mir-543 MIR-543







hsa-mir-548d-1 MIR-548D-1

hsa-mir-548d-2 MIR-548D-2


hsa-mir-554 MIR-554

hsa-mir-555 MIR-555

hsa-mir-568 MIR-568


hsa-mir-569 MIR-569

hsa-mir-577 MIR-577

hsa-mir-579 MIR-579

hsa-mir-580 MIR-580

hsa-mir-581 MIR-581

hsa-mir-582 MIR-582


hsa-mir-584 MIR-584

hsa-mir-585 MIR-585

hsa-mir-591 MIR-591

hsa-mir-592 MIR-592


hsa-mir-595 MIR-595

hsa-mir-596 MIR-596

hsa-mir-597 MIR-597

hsa-mir-598 MIR-598


hsa-mir-599 MIR-599


hsa-mir-542 MIR-542








hsa-mir-491 MIR-491







hsa-mir-499 MIR-499



hsa-mir-503 MIR-503


hsa-mir-504 MIR-504


hsa-mir-505 MIR-505


hsa-mir-506 MIR-506

hsa-mir-508 MIR-508

hsa-mir-509-1 MIR-509-1

hsa-mir-514-1 MIR-514-1

hsa-mir-514-2 MIR-514-2

hsa-mir-520f MIR-520F

hsa-mir-525 MIR-525




hsa-mir-541 MIR-541







79






mmu-mir-669h MMU-MIR-669H

mmu-mir-669j MMU-MIR-669J

mmu-mir-669k MMU-MIR-669K


hsa-mir-671 MIR-671







































hsa-mir-744 MIR-744


hsa-mir-758 MIR-758






hsa-mir-766 MIR-766

hsa-mir-767 MIR-767

hsa-mir-770 MIR-770


hsa-mir-802 MIR-802






hsa-mir-874 MIR-874




hsa-mir-708 MIR-708








mmu-mir-883A MMU-MIR-883A

mmu-mir-883B MMU-MIR-883B

hsa-mir-885 MIR-885

hsa-mir-889 MIR-889



hsa-mir-920 MIR-920

hsa-mir-921 MIR-921

hsa-mir-922 MIR-922

hsa-mir-923 MIR-923

hsa-mir-924 MIR-924

hsa-mir-933 MIR-933

hsa-mir-934 MIR-934

hsa-mir-935 MIR-935

hsa-mir-936 MIR-936

hsa-mir-937 MIR-937

hsa-mir-938 MIR-938

hsa-mir-940 MIR-940

hsa-mir-941 MIR-941

hsa-mir-942 MIR-942








hsa-mir-876 MIR-876


hsa-mir-877 MIR-877


80

Expression, Control, Reporter Vectors

miRNASelect™ Expression and Control Vectors

MICRORNA ANALYSIS


miRNASelect™ pEGP-mir Cloning and Expression Vector 100 µL MIR-EXP-GP-C

miRNASelect™ pEP-mir Cloning and Expression Vector 100 µL MIR-EXP-C

Our miRNASelect™ Mammalian Expression Vectors provide an easy, efficient method to clone a miRNA precursor from any species. The desired miRNA sequence is cloned into a human ß-globin intron contained within the vector. Two vector formats are available: The pEP vector contains a puromycin selection

marker The pEGP vector contains a GFP-puromycin fu-

sion to allow selection by either marker Each expression vector is provided with a null (empty) control vector at no extra charge.


Recent Product Citations 1. Esposito, F. et al. (2012). Down-regulation of the miR-25 and miR-30d contributes to the development of anaplastic thyroid carcinoma

targeting the polycomb protein EZH2. J. Clin. Endocrinol. Metab. 97:E710-E718. (MIR-EXP-GP-C) 2. Yuan, L. et al. (2012). MicroRNA-212 displays tumor-promoting properties in on-small cell lung cancer cells and targets the hedgehog

pathway receptor PTCH1. Mol. Biol. Cell 23:1423-1434. (MIR-EXP-GP-C)

miRNASelect™ Null Control Vectors


miRNASelect™ pEGP-mir Null Control Vector 100 µL MIR-NULL-GP

miRNASelect™ pEP-mir Null Control Vector 100 µL MIR-NULL

Our Null Control vectors have no cloned miRNA sequence. They are useful in conjunction with vectors from our miRNASelect™ Precursor Clone Collection.

Recent Product Citations 1. Dar, A. et al. (2013). The role of miR-18b in MDM2-p53 pathway signaling and melanoma progression. J. Natl. Cancer Inst. 105:433-442.

(MIR-NULL) 2. Larsen, M. et al. (2014). miRNA-130a regulates C/EBP-epsilon expression during granulopoiesis. Blood 123:1079-1089. (MIR-NULL-GP) 3. Yuan, L. et al. (2012). MicroRNA-212 displays tumor-promoting properties in on-small cell lung cancer cells and targets the hedgehog

pathway receptor PTCH1. Mol. Biol. Cell 23:1423-1434. (MIR-NULL-GP)




















MICRORNA ANALYSIS miRNA Viral Expression

81

RAPAd® miRNA Adenoviral Expression System


1 Kit VPK-253 RAPAd® miRNA Adenoviral Expression System

RAPAd® Adenoviral Expression Systems produce recombinant adenovirus with substantial reduction in wild-type adenovirus, while doing so in a much shorter 2 week time frame. The RAPAd® miRNA Adenoviral Expression System is specifically designed to deliver miRNA sequences into your target cell.

Virtually No Wild-Type Virus: Backbone vector engineered to produce <300 wild-type plaques per 109 particles, compared with 104-106 WT plaques per 109 VP with most other methods

Faster Virus Production: Virus generated in 2 weeks compared to a few months with traditional methods

For more information on our complete selection of RAPAd® Adenovirus Ex-pression Systems for gene expression

studies, please see page 49.


Recent Product Citation Li, P. et al. (2013). MicroRNA-638 is highly expressed in human vascular smooth muscle cells and inhibits PDGF-BB-induced cell proliferation and migration through targeting orphan nuclear re-ceptor NOR1. Cardiovasc. Res. 10.1093/cvr/cvt082. (VPK-253)

miRNA Retroviral Expression Vector


10 µg RTV-017 pMXs-miR-GFP/Puro Retroviral Vector

Our pMXs-miR-GFP/Puro Retroviral Vector allows you to clone a miRNA sequence of interest for pack-aging into a recombinant retrovirus for delivery into a target cell.

For efficient packaging of your miRNA into an MMLV-based retrovirus, use one of our Platinum Retroviral Packaging Cell Lines found on page 64.

Recent Product Citation Mansour, M. et al. (2013). The TAL1 complex targets the FBXW7 tumor suppressor by activating miR-223 in human T cell acute lymphoblastic leukemia. J. Exp. Med. 210:1545-1557. (RTV-017)

Adenovirus Production using the RAPAd® Adenoviral Expression System.

Reporter System, Knockdown Enhancer MICRORNA ANALYSIS

82

RNAiBoost™ Reagent Kit for miRNA and siRNA Enhancement


20 reactions RNAI-200

100 reactions RNAI-201 RNAiBoost™ Reagent Kit

RNA interference can occur in the presence of either siRNA or the mature form of miRNA. The RNABoost™ Reagent Kit increases the level of interference in the presence of siRNA. It also increases the rate of processing of pre-miRNA into mature miRNA.


miRNASelect™ Functional Analysis Reporter System


1 Kit MIR-GFP miRNASelect™ pMIR-GFP Reporter System

Assay Principle. If miRNA is present and binds to the 3’ UTR, translation of the GFP gene is repressed, resulting in loss of fluo-rescence.

The miRNASelect™ Functional Analysis Reporter System pro-vides a simple method for the evaluation of potential targets of miRNA. Binding of miRNA se-quences to their suspected tar-gets results in repression of trans-lation. In this system the miRNA target sequence, such as a 3’ UTR, is cloned into the provided pMIR-GFP vector in the multiple cloning sites immediately downstream of the GFP gene. If the miRNA is present and binds to the target sequence, translation is re-pressed and no green fluores-cence appears. If the miRNA does not bind the target se-quence, GFP translation occurs normally and green fluorescence may be seen.

OXIDATIVE STRESS / DAMAGE Oxidative Stress Overview

Measuring Oxidative Stress

84

Marker or

Type of Damage Sample Type

Cells Tissues Blood Urine Other

Protein Damage (p. 85-89)

Protein carbonyl content (PCC) X X X

3-Nitrotyrosine X X X

BPDE Protein Adduct X X X

Advanced Glycation End Products (AGE) X X X

Carboxyethyl Lynsine (CEL) X X X

Carboxymethyl Lynsine (CML) X X X

Methylglyoxal X X X

Advanced Oxidation Protein Products (AOPP) X X X

Lipid Peroxidation

(p. 90-93)

4-Hydroxynonenal (4-HNE) X X X

Malondialdehyde (MDA) X X X X

8-iso-Prostaglandin F2 (8-Isoprostane) X X X X

Oxidized Low Density Lipoprotein (OxLDL) X

DNA / RNA Damage and

Repair (p. 94-100)

8-hydroxyguanosine (8-OHG) X X X X Cerebrospinal Fluid

8-hydroxydeoxyguanosine (8-OHdG) X X X X

Abasic (AP) sites X X

Aldehyde DNA Damage (Etheno adducts) X X

BPDE DNA Adduct X X

Comet Assay X

Double-strand DNA breaks X

UV DNA Damage (CPD and 6-4PP) X

Reactive Oxygen Species

(p. 102-105)

Universal ROS X X X X

Hydrogen Peroxide X X X X

Nitric Oxide X X X X Saliva

Antioxidants & Antioxidant

Capacity (p. 106-110)

Superoxide Dismutase X X X X

Catalase X X X

Glutathione X X X X

Total Antioxidant Capacity (TAC) X X X X Food

Oxygen Radical Antioxidant Capacity (ORAC) X X X Food

Hydroxyl Radical Antioxidant Capacity (HORAC) X X X Food

Cellular Antioxidant Capacity (CAA) Antioxidant compounds

Oxidative stress may be measured using one of three primary methods:

Measure the reactive oxygen species (ROS) directly Measure the presence of antioxidants Measure the resulting damage to proteins, lipids, DNA or RNA (most reliable)

Use the following table to determine the best oxidative stress assays for your samples.


OXIDATIVE STRESS / DAMAGE Protein Damage

85

Assays and Reagents for Protein Damage

Cellular proteins are subject to damage in the presence of reactive oxygen species (ROS). The resulting protein damage may take the form of nitration or oxidation of various amino acid residues, or may result in formation of advanced glycation end products (AGE) or ad-vanced oxidation protein products (AOPP). We have developed unique assays to detect protein damage with higher sensitivity and more user-friendly protocols.

OxiSelect™ Nitrotyrosine Assay Kits and Antibodies


Nitrotyrosine ELISA Kit Colorimetric 96 Assays STA-305

5 x 96 Assays STA-305-5

Nitrotyrosine Immunoblot Kit Immunoblot/ECL 10 Blots STA-303

Goat Anti-Nitrotyrosine Polyclonal Antibody Immunoblot/ELISA 100 µg STA-003

Rabbit Anti-Nitrotyrosine Polyclonal Antibody Immunoblot/ELISA 100 µg STA-004

Protein Tyrosine Nitration Control (Nitrotyrosine-BSA) Immunoblot/ECL 10 µg STA-304

Formation of 3-Nitrotyrosine During Oxidative Stress.

Recent Product Citations 1. Toth, P. et al. (2014). Resveratrol treatment rescues neurovas-

cular coupling in aged mice: role of improved cerebromicrovas-cular endothelial function and downregulation of NADPH oxi-dase. Am. J. Physiol. Heart Circ. Physiol. 306:H299-H308. (STA-305)

2. Li, F.C. et al. (2013). Transition from oxidative stress to nitrosa-tive stress in rostral ventrolateral medulla underlies fatal intoxica-tion induced by organophosphate mevinphos. Toxicol. Sci. 135:202-217. (STA-305)

3. Medina, J.P. et al. (2013). Angiotensin receptor-mediated oxida-tive stress is associated with impaired cardiac redox signaling and mitochondrial function in insulin-resistant rats. Am. J. Physiol. Heart Circ. Physiol. 305:H599-H607. (STA-305)

4. Kong, X. et al. (2013). Pioglitazone enhances the blood pressure-lowering effect of losartan via synergistic attenuation of angio-tensin II-induced vasoconstriction. J. Renin Angiotensin Aldos-terone Syst. 10.1177/1170320313489061. (STA-305)

5. Lupachyk, S. et al. (2013). Endoplasmic reticulum stress plays a key role in the pathogenesis of diabetic peripheral neuropathy. Diabetes 62:944-952. (STA-305)

6. Cho, W.K. et al. (2013). IL-13 receptor 2-arginase 2 pathway mediates IL-13-induced pulmonary hypertension. Am. J. Physiol. Lung Cell Mol. Physiol. 304:L112-L124. (STA-305)

7. Montez, P. et al. (2012). Angiotensin receptor blockade recovers hepatic UCP2 expression and aconitase and SDH activities and ameliorates hepatic oxidative damage in insulin resistant rats. Endocrinology 153:5845-5856. (STA-305)

8. Tahrani, A.A. et al. (2012). Obstructive sleep apnea and diabetic neuropathy: a novel association in patients with type 2 diabetes. Am. J. Respir. Crit. Care Med. 186:434-441. (STA-305)

9. Suvakov, S. et al. (2012). Glutathione-S-transferase A1, M1, P1 and T1 null or low-activity genotypes are associated with en-hanced oxidative damage among haemodialysis patients. Nephrol. Dial. Transplant. 10.1093/ndt/gfs369. (STA-305)

10.Shevalye, H. et al. (2012). Metanx alleviates multiple manifesta-tions of peripheral neuropathy and increases intraepidermal nerve fiber density in Zucker diabetic fatty rats. Diabetes 61:2126-2133. (STA-305)

Our OxiSelect™ Nitrotyrosine Assay Kits provide a simple method to measure the formation of 3-nitrotyrosine in proteins. This assay is available in two formats: a 96-well competitive ELISA and an im-munoblot kit. The ELISA format can detect the pres-ence of 3-nitrotyrosine as low as 10 nM. Both kits can detect nitrotyrosine in protein from any species.


86


OxiSelect™ Protein Carbonyl Assay Kits


OxiSelect™ Protein Carbonyl ELISA Kit 96 Assays STA-310


OxiSelect™ Protein Carbonyl Spectrophotometric Assay Spectrophotometric 40 Assays STA-315

OxiSelect™ Protein Carbonyl Immunoblot Kit Immunoblot/ECL 10 Blots STA-308

Colorimetric

Oxidized Protein Immunoblot Control (Carbonyl-BSA) Immunoblot/ECL 10 µg STA-309

OxiSelect™ Protein Carbonyl Fluorometric Assay Fluorometric 100 Assays STA-307

Protein Carbonyl ELISA Kit Sensitive: Detects samples as low as 10

µg/ml Greater Sample Retention: No concentration

or TCA precipitation steps that contribute to sample loss

Assay Principle for the OxiSelect™ Protein Oxidation Immunoblot Kit (STA-308).

Protein Carbonyl Immunoblot Kit No Molecular Weight Shift: DNPH derivatization

after immunoblotting allows direct comparison of oxidized and non-oxidized protein fingerprints

The most common products of protein oxidation in biological samples are the carbonyl derivatives of Pro, Arg, Lys and Thr residues. Such derivatives are chemically stable and serve as markers for oxidative stress in most types of reactive oxygen species. Our OxiSelect™ Protein Carbonyl Assay Kits provide rapid, efficient methods for detection of protein car-bonyls. Four assay formats are available: im-munoblot, ELISA, fluorometric and spectrophotomet-ric. All formats are suitable for use with purified pro-tein, plasma, serum, or cell lysate samples from any species.

Recent Product Citations 1. Cui, Z. et al. (2014). Identification of the immunoproteasome as a

novel regulator of skeletal muscle differentiation. Mol. Cell Biol. 34:96-109. (STA-308)

2. Pons, D. et al. (2012). Initial activation status of the antioxidant response determines sensitivity to carboplatin/paclitaxel treatment of ovarian cancer. Anticancer Res. 32:4723-4728. (STA-308)

3. Cristovao, A.C. et al. (2012). NADPH oxidase 1 mediates -synucleinopathy in Parkinsons’ Disease. J. Neurosci. 32:14465-14477. (STA-308)

4. Bitar, M. et al. (2012). Decline in DJ-1 and decreased nuclear translocation of Nrf2 in Fuchs endothelial corneal dystrophy. In-vest. Ophthalmol. Vis. Sci. 53:5806-5813. (STA-308)

5. Cannizzo, E. et al. (2012). Age-related oxidative stress compro-mises endosomal proteostasis. Cell Report 2(1):136-149. (STA-308)

6. Satapati, S. et al. (2012). Elevated TCA cycle function in the pa-thology of diet-induced hepatic insulin resistance and fatty liver. J. Lipid Res. 53:1080-1092. (STA-308)

7. Vulusevic, B. et al. (2014). Glyoxalase-1 overexpression in bone marrow cells reverses defective neovascularization in STZ-induced diabetic mice. Cardiovasc. Res. 101:306-316. (STA-310)

8. Dai, D.F. et al. (2013). Global proteomics and pathway analysis of pressure-overload-induced heart failure and its attenuation by mitochondrial-targeted peptides. Circ. Heart Fail. 6:1067-1076. (STA-310)

9. Ungvari, Z. et al. (2013). Testing predictions of the oxidative stress hypothesis of aging using a novel invertebrate model of longevity: the giant clam (Tridacna derasa). J. Gerontol. A Biol. Sci. Med. Sci. 68:359-367. (STA-310)

10.Young, K. et al. (2013). Each to their own: skeletal muscles of different function use different biochemical strategies during aesti-vation at high temperature. J. Exp. Biol. 216:1012-1024. (STA-310)

11.Fishman , J. et al. (2012). Oxidative modification of the intestinal mucus layer is a critical but unrecognized component of trauma hemorrhagic shock-induced gut barrier failure. Am. J. Physiol. Gastrointest. Liver Physiol. 304:G57-G63. (STA-310)

12.Murakami, Y. et al. (2012). Receptor interacting protein kinase mediates necrotic cone but not rod cell death in a mouse model of inherited degeneration. PNAS 10.1073/pnas.1206937109. (STA-310)

13.Kang, K.A. et al. (2012). Baicalein inhibits oxidative stress-induced cellular damage via antioxidant effects. Toxic. And Ind. Health 28:412-421. (STA-310)

14.Gannon, A.M. et al. (2012). Cigarette smoke exposure leads to follicle loss via an alternative ovarian cell death pathway in a mouse model. Toxicol. Sci. 125:274-284. (STA-310)

15.Tang, H. et al. (2011). Intrinsic apoptosis in mechanically venti-lated human diaphragm: linkage to a novel Fox/FoxO1/Stat3-Bim axis. FASEB J. 25:2921-2936. (STA-310)

16.Robinson, C.K. et al. (2011). A major role for nonenzymatic anti-oxidant processes in the radioresistance of Halobacterium salina-rum. J. Bacteriol. 193:1653-1662. (STA-310)


87



OxiSelect™ AOPP Assay Kit

Advanced oxidation protein products are toxins cre-ated during oxidative stress in patients with diabetes mellitus, atherosclerosis, renal complications, and HIV. Our OxiSelect™ AOPP Assay Kit provides a quick, easy method for assessing AOPP levels.


OxiSelect™ AOPP Assay Kit Colorimetric 200 Assays STA-318

50 µL STA-319 AOPP-Human Serum Albumin N/A

Fast: Obtain results in <30 minutes Sensitive: Detect concentrations as low as 5 µM

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

Free HSA AOPP-HSA

OD

34

0n

m

Untreated Human Serum Albumin and AOPP-HSA Positive Control Tested with the OxiSelect™ AOPP Assay Kit.

OxiSelect™ BPDE Protein Adduct ELISA Kit

Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, cigarette smoke, and automotive exhaust. They may also be found in some cooked foods. One PAH, benzo(a)pyrene, was the first chemical carcinogen to be discovered. Through a series of enzymatic reac-tions, benzo(a)pyrene is converted to benzo(a)pyrene 7,8 diol-9,10 epoxide (BPDE) which attacks both proteins and DNA. Our OxiSelect™ BPDE Protein Adduct ELISA Kit provides a convenient method to measure the modification of proteins by BPDE.

Sensitive: Detect concentrations as low as 60 ng/mL

Convenient: Quantify on a standard microplate reader

Versatile: Suitable for use with cell lysates, tis-sue homogenates, plasma or serum


OxiSelect™ BPDE Protein Adduct ELISA Kit Colorimetric 96 Assays STA-301

BPDE-BSA Standard Curve Generated Using the OxiSelect™ BPDE Protein Adduct ELISA Kit.

For information on our BPDE DNA Adduct ELISA Kit, please see page 99.

Recent Product Citations 1. Bloomer, R. et al. (2013). Safety profile of caffeine and 1,3-

dimethylamylamine supplementation in healthy men. Human and Exp. Toxicol. 10.1177/0960327113475680. (STA-318)

2. Park, S.H. et al. (2012). Effects of neutral pH and low-glucose degradation product-containing peritoneal dialysis fluid on sys-temic markers of inflammation and endothelial dysfunction: a randomized controlled 1-year follow-up study. Nephrol. Dial. Transp. 27:1191-1199. (STA-318)

3. Anderson, D. et al. (2010). Albumin-based microbubbles bind up-regulated scavenger receptors following vascular injury. J. Biol. Chem. 285:40645-40653. (STA-318)



Advanced glycation end products (AGE) are formed during the Maillard reaction where reducing carbohy-drates react with lysine side chains and N-terminal amino groups of various macromolecules, particularly proteins. These AGE products can adversely affect the function of the affected proteins and play a role in atherosclerosis, diabetes, aging and renal disease. Our OxiSelect™ Advanced Glycation End Product Kits are designed for the rapid detection of AGE pro-tein adducts. We offer assays to study generic AGE formation or specific AGE strucutres including N-(Carboxyethyl) lysine (CEL), N-(Carboxymethyl) lysine (CML), and methylglyoxal (MG). All kits will detect AGE structures from protein of any species.

Sensitive: Detect levels as low as 1 µg/mL of AGE-protein adduct

Versatile: Compatible with cell lysates, plasma, serum, or purified proteins

Advanced Glycation End Products (AGE) Pathways.

OxiSelect™ Advanced Glycation End Product Kits & Antibodies


Colorimetric 96 Assays STA-817


Glycoaldehyde-AGE-BSA N/A 100 µg STA-348

OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit

OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit

Our OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit detects a variety of AGE structures including CML and pentosidine. It does not detect CEL or methylglyoxal (MG). Samples are added to a plate coated with an AGE-protein conjugate. AGE-protein adducts in the sample compete with the AGE-coated plate for antibody bind-ing. High AGE adduct content in a sample results in less binding of the antibody to the plate, producing a low signal.


OxiSelect™ N-(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit Colorimetric 96 Assays STA-813

CEL-BSA N/A 100 µg STA-302

OxiSelect™ N-(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit

Sensitive: Detect levels as low as 100 ng/mL of CEL-protein adduct


The OxiSelect™ N-(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit detects CEL protein adducts in a variety of samples including cell lysates, blood sam-ples, and other protein sources.

OxiSelect™ AGE Competitive ELISA Kit Standard Curve.




OxiSelect™ Methylglyoxal (MG) Competitive ELISA Kit 96 Assays STA-811


Mouse Anti-Methylglyoxal Monoclonal Antibody Immunoblot/

Immunohistochemistry 100 µg STA-011

MG-BSA N/A 100 µg STA-306

Colorimetric

OxiSelect™ N-(Carboxymethyl) Lysine (CML) Assays and Antibodies

Oxidized / Nitrated Proteins


Copper (Cu++) Oxidized Human Low Density Lipoprotein (LDL) 100 µg STA-214

Malondialdehyde (MDA) Modified Human Albumin 100 µg STA-210

Malondialdehyde (MDA) Modified Human Apolipoprotein B-100 100 µg STA-211

Malondialdehyde (MDA) Modified Human Low Density Lipoprotein (LDL) 100 µg STA-212

Nitrated Human Low Density Lipoprotein (LDL) 100 µg STA-213

All proteins are provided at a concentration of 1.0 mg/mL.




OxiSelect™ N-(Carboxymethyl) Lysine (CML) Immunoblot Kit Immunoblot 10 Blots STA-313

Goat Anti-N-CML Polyclonal Antibody Immunoblot/ELISA 100 µg STA-013

Rabbit Anti-N-CML Polyclonal Antibody Immunoblot/ELISA 100 µg STA-014

CML-BSA Control N/A 100 µg STA-314

OxiSelect™ N-(Carboxymethyl) Lysine (CML) Competitive ELISA Kit

OxiSelect™ Methylglyoxal (MG) Assays and Antibodies

Sensitive: Detect levels as low as 200 ng/mL of MG-protein adduct


The OxiSelect™ Methylglyoxal (MG) Competitive ELISA Kit detects MG protein adducts in a variety of samples including cell lysates, blood samples, and other protein sources.

Sensitive: Detect levels as low as 3 ng/mL of CML-protein adduct with the ELISA kit


The OxiSelect™ N-(Carboxymethyl) Lysine (CML)Competitive ELISA Kit detects CML protein adducts in a variety of samples including cell lysates, blood samples, and other protein sources.

CML Protein Adduct Detected in Human Plasma with the OxiSelect™ CML Competitive ELISA Kit.

Assays and Reagents for Lipid Peroxidation

Lipid peroxidation is a well-defined mechanism of cellular damage in both animals and plants that occurs during aging and in some disease states. Our OxiSelect™ Lipid Peroxi-dation Assays allow you to quickly and easily quantify the most common markers and by-products of lipid peroxidation.

OxiSelect™ TBARS Assay Kit

The TBARS assay is a well-established method for screening and monitoring lipid peroxidation via the by-product malondialdehyde (MDA). MDA forms a 1:2 adduct with thiobarbituric acid. Our OxiSelect™ TBARS Assay Kit provides a more user-friendly protocol for quantitation of the MDA-TBA adduct compared to other commercial assays. This assay detects total MDA, both free and in pro-tein adducts, in a variety of samples including cell and tissue lysates, plasma, and urine.


Colorimetric or Fluorometric

200 Assays STA-330

5 x 200 Assays STA-330-5 OxiSelect™ TBARS Assay Kit (MDA Quantitation)

Fast: Obtain results in 30 minutes Sensitive: Smaller reaction volumes require less

sample; detect as little as 2 µM Convenient: 96-well format; no glass tubes are

required

90

OXIDATIVE STRESS / DAMAGE Lipid Peroxidation

Recent Product Citations 1. Chang, Q. et al. (2014). Cytochrome P450 2C epoxygenases

mediate photochemical stress-induced death of photoreceptors. J. Biol. Chem. 289:8337-8352.

2. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adap-tation. J. Gerontol. A Biol. Sci. Med. Sci. 10.1093/gerona/glt122.

3. Fu, L. et al. (2012). Ethyl pyruvate reduces ventilation-induced neutrophil infiltration and oxidative stress. J. Lipid Res. 53:1080-1092.

4. Brindeiro, C.M. et al. (2012). Tempol prevents altered K+ chan-nel regulation of afferent arteriolar tone in diabetic rat kidney. Hypertension 59:657-664.

5. Joshi, S.G. et al. (2011). Nonthermal dielectric-barrier discharge plasma-induced inactivation involves oxidative DNA damage and membrane lipid peroxidation in Escherichia coli. Antimicrob. Agents Chemother. 55:1053-1062.

6. Kasaikina, M.V. et al. (2011). Roles of the 15-kDa Selenoprotein (Sep15) in redox homeostasis and cataract development re-vealed by the analysis of Sep15 knockout mice. J. Biol. Chem. 286:33203-33212.

7. Fedeles, B.I. et al. (2011). Chemical genetics analysis of an aniline mustard anticancer reveals complex I of the electron transport chain as a target. J. Biol. Chem. 286:33910-33920.

8. Wojciechowski, P. et al. (2010). Resveratrol arrests and re-gresses the development of pressure overload- but not volume overload-induced cardiac hypertrophy in rats. J. Nutr. 140:962-968.


OxiSelect™ MDA (Malondialdehyde) Assays and Antibodies

As a common by-product of lipid peroxidation, malondialdehyde (MDA) is a well-accepted marker of oxidative stress. Modification of proteins by MDA can cause structural and functional changes in oxidized proteins. We offer assays and antibodies to measure MDA in a variety of formats. Kits are available to measure total MDA as well as MDA protein adducts specifically.

Note: MDA is most reliably detected in fresh sam-ples, or in samples that have been frozen for a maxi-mum of 1-2 months. For samples stored for longer periods, consider testing other markers of lipid peroxi-dation such as 4-HNE or 8-isoprostane.

MDA-TBA Standard Curve Using a Standard Plate Reader.

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0 10 20 30 40

OD

54

0n

m

MDA (µM)

Colorimetric Assay

91



OxiSelect™ MDA Adduct Assay Kits


Goat Anti-Malondialdehyde (MDA) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-031

Rabbit Anti-Malondialdehyde (MDA) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-032

Recent Product Citations 1. Montez, P. et al. (2012). Angiotensin receptor blockade recov-

ers hepatic UCP2 expression and aconitase and SDH activities and ameliorates hepatic oxidative damage in insulin resistant rats. Endocrinology 153:5845-5856. (STA-331)

2. Lazrak, A. et al. (2011). Regulation of alveolar epithelial Na+ channels by ERK1/2 in chlorine breathing mice. Am. J. Respir. Cell Mol. Biol. 10.1165/rcmb.2011-0309OC. (STA-331)

3. Zarogiannis, S.G. et al. (2011). Ascorbate and deferoxamine administration post chlorine exposure decrease mortality and lung injury in mice. Am. J. Respir. Cell Mol. Biol. 45:386-392. (STA-331)

4. Hall, J.A. et al. (2010). Absence of thyroid hormone activation during development underlies a permanent defect in adaptive thermogenesis. Endrocrinology 151:4573-4582. (STA-331)

5. Barabutis, N. et al. (2008). Antioxidant activity of growth hor-mone-releasing hormone antagonists in LNCaP human pros-tate cancer cell line. PNAS 105:20470-20475. (STA-331)

Sensitive: ELISA kit detects MDA protein ad-ducts as low as 6 pmol/mL

Versatile: Suitable for use with serum, plasma, cell lysates or tissue homogenates

Immunoblot of MDA-BSA Control Using the OxiSelect™ MDA Immunoblot Kit. Immunoblot control was electroblotted onto a nitrocellulose membrane, followed by detection with the provided anti-MDA antibody.


OxiSelect™ MDA Immunoblot Kit Immunoblot 10 Blots STA-331

OxiSelect™ MDA Adduct Competitive ELISA Kit Colorimetric 96 Assays STA-832


MDA-BSA N/A 100 µg STA-333

OxiSelect™ MDA Polyclonal Antibodies

Our MDA Adduct Assay Kits provide simple methods to measuring these protein adducts in a variety of sample types. The MDA Adduct Competitive ELISA Kit is a sensitive method for the quantitation of MDA in proteins from cells, tissues, or blood. Samples are added to a malondialdehyde protein conjugate-coated plate. The MDA in the sample competes with the MDA on the plate for binding to the primary anti-MDA antibody. A high concentration of MDA in the sample results in little to no antibody binding to the plate, producing a low signal. Our MDA Immunoblot is a convenient method for qualitative measurement of MDA protein adducts.



Standard Curve Generated with the OxiSelect™ HNE Adduct Competitive ELISA Kit.

OxiSelect™ HNE ELISA Kits and Antibodies


OxiSelect™ HNE Adduct Competitive ELISA Kit Colorimetric 96 Assays STA-838


Goat Anti-4-Hydroxynonenal (HNE) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-034

Rabbit Anti-4-Hydroxynonenal (HNE) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-035

HNE-BSA N/A 100 µg STA-335

Sensitive: ELISA kit detects protein adducts as low as 2 µg/mL

Versatile: Suitable for use with serum, plasma, cell lysates or tissue homogenates

4-hydroxynonenal (4-HNE) is a well-known by-product of lipid peroxidation and is widely accepted as a stable marker for oxidative stress. HNE protein adducts are typically stable when frozen for up to 6 months or more. Our OxiSelect™ HNE Adduct Competitive ELISA Kit provides a simple, user-friendly way to assess HNE adduct formation on lysine, histidine and/or cysteine. Our polyclonal antibodies against HNE recognize HNE adducts to lysine, histidine, and/or cysteine and are suitable for use with Western Blot or ELISA ap-plications.

OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit (8-isoprostane)


OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit 96 Assays STA-337

5 x 96 Assays STA-337-5 Colorimetric

8-iso-Prostaglandin F2 is produced in membrane phospholipids and has been implicated in athero-genesis, rheumatoid arthritis and carcinogenesis. The OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit provides rapid, sensitive detection of 8-iso-PGF2as low as 50 pg/mL. The assay is suitable for quantita-tion of 8-isoprostane in a variety of sample types in-cluding cell and tissue lysates, plasma, serum, and urine.

Recent Product Citations 1. Dugas, T.R. et al. (2014). Hydrogen sulfide cytoprotective sig-

naling is endothelial nitric oxide synthase-nitric oxide dependent. PNAS 111:3182-3187.

2. Karakus, E. et al. (2013). Agomelatine: an antidepressant with new potent hepatoprotective effects on paracetamol-induced liver damage in rats. Human and Experimental Toxicol. 10.0177/0960327112472994.

3. Mollo, R. et al. (2012). Effect of -lipoic acid on platelet reactivity in type 1 diabetic patients. Diabetes Care 35:196-197.

4. Thompson, C.M. et al. (2012). Comparison of the effects of hexavalent chromium in the alimentary canal of F344 rats and B6C3F1 mice following exposure in drinking water: implications for carcinogenic modes of action. Toxicol. Sci. 125:79-90.



OxiSelect™ Human Oxidized LDL ELISA Kits

LDL contains a hydrophobic core of various lipids surrounded by one molecule of Apolipoprotein B-100 (ApoB-100), which promotes solubility of the LDL in blood. LDL, often described as “bad” cholesterol, is even more dangerous when it becomes oxidized. Oxi-dized LDL (OxLDL) is more reactive with surrounding tissues and can collect within the inner lining of arter-ies. Our OxiSelect™ Human Oxidized LDL ELISA Kits are designed for the detection and quantitation of modi-fied LDL in human plasma or serum. Kits are avail-able to detect MDA-LDL, CML-LDL, or HNE-LDL in either the protein or lipid component of LDL. Our OxPL-LDL kit specifically detects oxidation in the phospholipid component of LDL.


OxiSelect™ Human Oxidized LDL ELISA Kit (CML-LDL Quantitation) Colorimetric 96 Assays STA-388

OxiSelect™ Human Oxidized LDL ELISA Kit (HNE-LDL Quantitation) Colorimetric 96 Assays STA-389

OxiSelect™ Human Oxidized LDL ELISA Kit (MDA-LDL Quantitation) Colorimetric 96 Assays STA-369

OxiSelect™ Human Oxidized LDL ELISA Kit (OxPL-LDL Quantitation) Colorimetric 96 Assays STA-358

Sensitive: Detect as little as 50 ng/mL of MDA-LDL, 150 ng/mL of CML-LDL, 150 ng/mL of HNE-LDL, or 100 ng/mL of OxPL-LDL

Quantitative: Compare unknown samples with provided copper oxidized LDL standard

Quantitation of MDA-LDL in Serum and Plasma Samples. Se-rum and plasma samples were treated with LDL Precipitation Solu-tion. Precipitated LDL pellets were resuspended in 1.6 mL of PBS before further diluting 1:160 in Assay Diluent according to the As-say Protocol. OxiSelect™ Human Oxidized LDL ELISA Assay Principle.

MDA is the most commonly found damage marker in oxidized LDL, but it can degrade in frozen samples after 1-2 months. CML and HNE, while less commonly found in OxLDL, may be more reliably detectable in samples that have been frozen for several months.

94

Assays for DNA & RNA Damage and Repair

DNA is arguably the most biologically significant target of oxidative and cellular stress. Continuous DNA damage has been implicated in age-related development of various can-cers. More recently, RNA damage has been described in conjunction with various neuro-logical diseases including Alzheimer’s and Parkinson’s diseases. We offer a wide range of assays to measure the most common types of DNA and RNA damage in cells.

OxiSelect™ Oxidative DNA Damage ELISA Kit (8-OHdG Quantitation)

Among the various types of oxidative DNA damage, 8-hydroxydeoxyguanosine (8-OHdG) is a ubiquitous marker of oxidative stress. 8-OHdG, one of the by-products of DNA oxidative damage, is physiologi-cally formed and enhanced by chemical carcino-gens. Our OxiSelect™ Oxidative DNA Damage ELISA Kit provides a powerful method for rapid quantitation of 8-OHdG in DNA samples. 8-OHdG is easily detect-able directly in serum and urine samples, after it is excised and secreted during the DNA repair proc-ess. It also may be detected in DNA extracted from cells or tissues of any species, following full DNA digestion into single bases.

Highly Sensitive: Detect as little as 100 pg/mL of 8-OHdG

Versatile: Suitable for use with urine, serum, and DNA extracted from cells or tissues


OxiSelect™ Oxidative DNA Damage ELISA Kit (8-OHdG Quantitation) Colorimetric 96 Assays STA-320


Recent Product Citations 1. Moore, J. et al. (2013). Protection of corneal epithelial stem cells

prevents ultraviolet A damage during corneal collagen cross-linking treatment for keratoconus. Br. J. Ophthalmol. 10.1136/bjophthalmol-2013-303816.

2. Kalghatgi, S. et al. (2013). Disruption of cell-cell junctions and induction of pathological cytokines in the retinal pigment epithe-lium of light-exposed mice. Sci. Transl. Med. 5:192ra85.

3. James, M.L. et al. (2013). VARA attenuates hyperoxia-induced impaired alveolar development and lung function in newborn mice. Am. J. Physiol. Lung Cell Mol. Physiol. 304:L803-L812.

4. Ravikumar, P. et al. (2013). Klotho protects lung epithelial cells against oxidative DNA damage. FASEB J. 27:722-723.

5. Singh, B. et al. (2013). MicroRNA-93 regulates NRF2 expression and is associated with breast carcinogenesis. Carcinogenesis 10.1093-carcin/bgt026.

6. Singh, B. et al. (2012). Superoxide dismutase 3 is induced by antioxidants, inhibits oxidative DNA damage and is associated with inhibition of estrogen-induced breast cancer. Carcinogene-sis 33:2601.2610.

7. Mercer, J.R. et al. (2012). The methyl xanthine caffeine inhibits DNA damage signaling and reactive species and reduces atherosclerosis in ApoE-/- mice. Arterioscler. Thromb. Vasc. Biol. 32:2461-2467.

8. Schutt, F. et al. (2012). Moderately reduced ATP levels promote oxidative stress and debilitate autophagic and phagocytic ca-pacities in human RPE cells. Invest. Ophthalmol. Vis. Sci. 53:5354-5361.

9. Tanabe, K. et al. (2012). Nicorandil as a novel therapy for ad-vanced diabetic nephrophathy in the eNOS-deficient mouse. Am. J. Physiol. Renal Physiol. 302:F1151-F1160.

10.Tzortzaki, E.G. et al. (2012). Oxidative DNA damage and so-matic mutations: a link to the molecular pathogenesis of chronic inflammatory airway diseases. Chest 141:1243-1250.

11.Xu, X. et al. (2012). Reactive oxygen species-triggered tro-phoblast apoptosis is initiated by endoplasmic reticulum stress via activation of caspase-12, CHOP, and the JNK pathway in Toxoplasma gondii infection in mice. Infect. Immun. 80:2121-2132.

12.Burnham, E. L. (2012). Protandim does not influence alveolar epithelial permeability or intrapulmonary oxidative stress in hu-man subjects with alcohol use disorders. Am. J. Physiol. Lung Cell Mol. Physiol. 302:L688-L699.

13.Pialoux, V. et al. (2011). Losartan abolishes oxidative stress induced by intermittent hypoxia in humans. J. Physiol. 589:5529-5537.

0

0.5

1

1.5

2

2.5

Negat

ive

Contro

l

1/20

dilu

tion

1/40

dilu

tion

1/80

dilu

tion

1/16

0 d

ilutio

n

1/32

0 d

ilutio

n

1/64

0 d

ilutio

n

OD

45

0 n

m

8-OHdG Levels in a Human Urine Sample.

OXIDATIVE STRESS / DAMAGE DNA / RNA Damage & Repair


95



OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation)

Similarly to 8-hydroxydeoxyguanosine (8-OHdG) forming during DNA oxidation, RNA can become oxidized resulting in 8-hydroxyguanosine (8-OHG). Oxidation of RNA has been implicated in a number of neurological diseases including Alzheimer’s and Parkinson’s diseases. Our OxiSelect™ Oxidative RNA Damage ELISA Kit provides a powerful method for rapid quantitation of 8-OHG in urine, serum or cerebrospinal fluid. It also may be used to detect 8-OHG in RNA extracted from cells or tissues of any species.

Highly Sensitive: Detect as little as 150 pg/mL of 8-OHG

Versatile: Suitable for use with urine, serum, cerebrospinal fluid, and DNA extracted from cells or tissues


OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation) Colorimetric 96 Assays STA-325


Recent Product Citations 1. Kannan, S. et al. (2012). Dendrimer-based postnatal therapy for

neuroinflammation and cerebral palsy in a rabbit model. Sci. Transl. Med. 4:130ra46.

2. Bazin, J. et al. (2011). Targeted mRNA oxidation regulates sun-flower seed dormancy alleviation during dry after-ripening. Plant Cell 23:2196-2208.

OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine Quantitation)


OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine Quantitation) Colorimetric

96 Assays STA-825


Various reactive nitrogen species (RNS) including peroxynitrite and nitrogen oxides can form during pathophysiological conditions. These RNS can ni-trate guanine bases to form 8-nitroguanine in both DNA and RNA. Nitrosative damage to DNA and RNA is a significant contributor to the age-related development of major inflammation-related diseases as well as colon, breast, and prostate cancers. Our OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit provides a simple method for rapid quanti-tation of 8-nitroguanine in urine, serum or plasma samples. The assay measures total 8-nitroguanine from both DNA and RNA combined.

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

0.01 0.1 1 10 100

8-OHG (ng/mL)O

D 4

50

nm

Standard Curve Generated with the OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine).

Standard Curve Generated with the OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG).



OxiSelect™ Oxidative DNA Damage Quantitation Kit (AP Sites)


OxiSelect™ Oxidative DNA Damage Quantitation Kit (AP Sites) Colorimetric 50 Assays STA-324

Highly Sensitive: Detect as few as 4-40 AP sites in 105 bp of DNA

Versatile: Suitable for use with genomic DNA from cells or tissues

Quantitative: Kit includes both oxidized and reduced DNA standards for absolute quantita-tion

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

0 10 20 30 40 50

AP Sites per 100,000bp

OD

45

0 n

m

Standard Curve Generated with the OxiSelect™ Oxidative DNA Damage Quantitation Kit (STA-324).

OxiSelect™ DNA Double-Strand Break Assay


OxiSelect™ DNA Double-Strand Break Staining Kit Immuno-

fluorescence 100 Assays STA-321

Double-strand breaks (DSB) are among the most dangerous types of DNA damage within cells. An early cellular response is phosphorylation of the his-tone variant H2AX at the site of the DSB. This trig-gers a cascade of events and appears to play a role in recruitment of repair factors to the damaged sites. Our OxiSelect™ DNA Double-Strand Break Staining Kit provides an easy-to-use method for detecting DNA breaks. The kit utilizes simple immunofluores-cence staining of the phosphorylated histone H2AX.

Fast: See staining results in about 3 hours Positive Control: DNA Double-strand break

inducer included in kit

DNA Double-Strand Break Formation in A549 Cells. A549 cells were seeded at 50,000 cells/well overnight. Immunofluores-cence staining was then performed according to the assay pro-tocol. (A) Untreated cells. (B) Cells treated with 100 µM eto-poside for one hour.

Oxidative DNA Damage can manifest in the forma-tion of apurinic or apyrimidinic (AP) sites, also known as loss of bases. Spontaneous base loss, if unrepaired, can inhibit transcription and may be mutagenic. Our OxiSelect™ Oxidative DNA Damage Quantita-tion Kit provides a simple, user-friendly method for measuring AP sites in DNA. The assay uses an al-dehyde reactive probe (ARP) which specifically re-acts with an aldehyde group on the open ring of the AP site, followed by labeling with Biotin and subse-quent detection by Streptavidin-enzyme conjugate.

Recent Product Citations 1. Messaoudi, N. et al. (2013). Global stress response in a pro-

karyotic model of DJ-1-associated Parkinsonism. J. Bacteriol. 195:1167-1178.

2. Zaika, E. et al. (2011). P73 protein regulates DNA damage repair. FASEB J. 25:4406-4414.



OxiSelect™ Comet Assays (Single Cell Gel Electrophoresis)


OxiSelect™ 3-Well Comet Assay Kit Light Microscopy

15 Wells STA-350

75 Wells STA-351

5 x 75 Wells STA-351-5

OxiSelect™ 3-Well Comet Assay Slides Light Microscopy

5 Slides STA-352

25 Slides STA-353

125 Slides STA-353-5

OxiSelect™ 96-Well Comet Assay Kit Light Microscopy 96 Wells STA-355

5 x 96 Wells STA-355-5

OxiSelect™ 96-Well Comet Assay Slides 1 Slide STA-356

5 Slides STA-356-5

OxiSelect™ Comet Assay Control Cells (includes positive and negative controls) N/A 1 Set STA-354

Light Microscopy

DNA damage can result from a variety of intracellu-lar and extracellular stimuli, and can manifest in a variety of mutations to the DNA including base modi-fications, missing bases and single-stranded or dou-ble-stranded breaks. Traditionally the comet assay, or single cell gel elec-trophoresis (SCGE), has been used as a well-published, high-level screening tool to measure DNA damage in single cells. Our OxiSelect™ Comet Assay Kits provide a quick, easy method to screen for DNA damage at a macro level. Our OxiSelect™ Comet Assay Slides have been specially treated for adhesion of low-melting agarose used in the assay. Damaged DNA moves farther in electrophoresis than intact DNA, causing a “tail” to form upon visualization under a fluorescence microscope.

Etoposide Treatment of Jurkat Cells. Jurkat cells were either untreated (left) or treated with etoposide (right) prior to perform-ing the OxiSelect™ Comet Assay.

Assay Principle for the OxiSelect™ Comet Assay Kit.

Recent Product Citations 1. Luo, Y. et al. (2013). SMC1-mediated intra-S-phase arrest

facilitates bocavirus DNA replication. J. Virol. 87:4017-4032. (STA-350, STA-351)

2. Li, Y.J. et al. (2012). Gold nanoparticles as a platform for creating a multivalent poly-SUMO chain inhibitor that also augments ionizing radiation. PNAS 109:4092-4097. (STA-350, STA-351)

3. Robin, T.P. et al. (2012). EWS/FLI1 regulates EYA3 in Ewing Sarcoma via modulation of miRNA-708, resulting in increased cell survival and chemoresistance. Mol. Cancer Res. 10:1098-1108. (STA-350, STA-351)

4. Choi, S.K. et al. (2012). Poly(ADP-ribose) polymerase 1 inhi-bition improves coronary arteriole function in type 2 diabetes mellitus. Hypertension 59:1060-1068. (STA-350, STA-351)

5. Tyagi, A. et al. (2011). Resveratrol selectively induces DNA damage, independent of Smad4 expression, in its efficacy against human head and neck squamous cell carcinoma. Clin. Cancer Res. 17:5402-5411. (STA-355)

98



OxiSelect™ UV-Induced DNA Damage Assays


OxiSelect™ UV-Induced DNA Damage ELISA Combo Kit (CPD/6-4PP) Colorimetric 96 Assays STA-322-C

OxiSelect™ UV-Induced DNA Damage ELISA Kit (CPD Quantitation) Colorimetric 96 Assays STA-322



5 x 96 Assays STA-323-5 OxiSelect™ UV-Induced DNA Damage ELISA Kit (6-4PP Quantitation)




OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kit (6-4PP) Colorimetric 96 Assays STA-328

OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kit (CPD)

OxiSelect™ UV-Induced DNA Damage ELISA Kits, for extracted DNA

Absorption of ultraviolet radiation can damage DNA by the formation of pyrimidine dimers. The two most common forms of pyrimidine dimers are cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) pyrimi-done photoproducts (6-4PP). Our OxiSelect™ UV-Induced DNA Damage Assays conveniently measure the formation of either CPD or 6-4PP in intact cells. Kits for each marker are avail-able in three formats: an ELISA for DNA extracted from cells or tissues, a Cell-Based ELISA, and a Cel-lular Immunostaining kit.

UV-Induced DNA Damage in HeLa Cells Treated with Ultravio-let Light for 30 Minutes and Visualized with the OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (CPD).

UV-Induced DNA Damage in HeLa Cells as Detected with the OxiSelect™ Cellular UV-Induced DNA Damage ELISA (CPD).

Recent Product Citations 1. Zirkin, S. et al. (2013). The PIM-2 kinase is an essential com-

ponent of the ultraviolet damage response that acts upstream to E2F-1 and ATM. J. Biol. Chem. 288:21770-21789. (STA-322)

2. Burgess, H.M. et al. (2011). Nuclear relocalisation of cytoplas-mic poly(A)-binding proteins PABP1 and PABP4 in response to UV irradiation reveals mRNA-dependent export of meta-zoan PABPs. J. Cell Sci. 124:3344-3355. (STA-322)

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

SecondaryAntibody Alone

UV Treated UntreatedO

D 4

50

nm


OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (CPD) Fluorescence Microscopy

96 Assays STA-327

OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (6-4PP) Fluorescence Microscopy

96 Assays STA-329

OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kits, for intact cells

OxiSelect™ Cellular UV-Induced DNA Damage Staining Kits, for intact cells

99



OxiSelect™ BPDE DNA Adduct ELISA Kit

Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, cigarette smoke, and automotive exhaust. One PAH, benzo(a)pyrene, was the first chemical carcinogen to be discovered. Through a series of enzymatic reac-tions, benzo(a)pyrene is converted to benzo(a)pyrene 7,8 diol-9,10 epoxide (BPDE) which attacks both pro-teins and DNA. Our OxiSelect™ BPDE DNA Adduct ELISA Kit pro-vides a convenient method to measure BPDE ad-ducts in DNA extracted from cells or tissues.

Sensitive: Detect concentrations as low as 30 ng/mL

Convenient: Quantify on a standard microplate reader


OxiSelect™ BPDE DNA Adduct ELISA Kit Colorimetric 96 Assays STA-357

BPDE-DNA Standard Curve Generated Using the OxiSelect™ BPDE DNA Adduct ELISA Kit.

For information on our BPDE Protein Adduct ELISA Kit, please see page 87.


OxiSelect™ Aldehyde-Induced DNA Damage ELISA Combo Kit (Ethenoadenosine / Ethenocytidine Quantitation)

Colorimetric 96 Assays STA-820-C

OxiSelect™ Aldehyde-Induced DNA Damage ELISA Kit (Ethenoadenosine Quantitation)


OxiSelect™ Aldehyde-Induced DNA Damage ELISA Kit (Ethenocytidine Quantitation)


OxiSelect™ Aldehyde-Induced DNA Damage Assays (Etheno Adducts)

Oxidation of phospholipids can lead to the formation of lipid hydroperoxides. These resulting short-lived hydrop-eroxides can either be converted to inert fatty acid alcohols, or can react with metals to form aldehydes such as malondialdehyde (MDA), 4-hydroxynonenal (HNE), acrolein, and crotonaldehyde. These aldehydes (which can also be formed through exposure to carcinogenic substances such as urethane or vinyl chloride) can damage DNA resulting in the formation of various etheno adducts, including 1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine. The presence of these bases can lead to base pair substitution mutations.

Our OxiSelect™ Aldehyde-Induced DNA Damage Assays conveniently measure the formation of either 1,N6-ethenodeoxyadenosine (ethenoadenosine) or 3,N4-ethenodeoxycytidine (ethenocytidine) in DNA extracted from cells or tissues. In addition, we offer a convenient combination kit that can measure both etheno bases in separate wells of the same plate.

Etheno Base Structures that Form Adducts with DNA During Oxidative Stress.

Recent Product Citation Chiu, C.Y. et al. (2014). Low-dose benzo(a)pyrene and its epox-ide metabolite inhibit myogenic differentiation in human skeletal muscle-derived progenitor cells. Toxicol. Sci. 10.1093/toxsci/kfu003.



Checkpoint Kinase Activity Assays


Checkpoint Kinase Activity Immunoblot Kit Immunoblot 20 Assays STA-413


5 x 96 Assays STA-414-5 96-Well Checkpoint Kinase Activity Assay Kit

Checkpoint kinases, specifically CHK1 and CHK2, are activated in response to DNA damage to subsequently phosphorylate Cdc25C prior to mitosis, which prompts cell cycle arrest. Mutation of these checkpoint kinases can ultimately lead to decreased DNA repair. Our Checkpoint Kinase Activity Assays allow you to conveniently measure the activity of CHK1 and CHK2. The assays use recombinant Cdc25C as a checkpoint kinase substrate. Phosphorylated Cdc25C (Ser216) is detected using a phospho-specific antibody. Checkpoint Kinase Activitiy Assays are available in two formats: a Western blot assay and a 96-well plate-based activity assay.


Global DNA Methylation ELISA Kit (5’-methyl-2’-deoxycytidine Quantitation)



Global DNA Methylation ELISA Kit

Our Global DNA Methylation and Hydroxymethyla-tion Assays provide a convenient, accurate way to quantify 5’-methyl-2’-deoxycytidine (5MedCyd) and 5-hydroxymethylcytosine respectively. Unknown samples are compared with a standard provided with each kit.

Sensitive: Detect as little as 15 nM of 5MedCyd Versatile: Suitable for use with any isolated

DNA as well as urine samples Convenient: Quantify on a standard microplate

reader

Methylated DNA Standard Curve Generated with the Global DNA Methylation ELISA Kit.

5MedCyd Levels in Human Urine Sample as Measured with the Global DNA Methylation ELISA Kit.

DNA methylation is an epigenetic change shown to be associated with nearly every biological process. In mammalian cells, DNA methylation is found predominantly at CpG dinucleotides; however, in certain cases such as embryonic stem cells it may also be found in non-CpG contexts. Due to the important role of DNA me-thylation in maintaining genomic stability, deregulation of DNA methylation is associated with various diseases including cancer.


OXIDATIVE STRESS / DAMAGE Hypoxia Assays

HIF-1 Alpha DNA Binding Activity Assay Kit

Detection Specificity of HIF-1 Alpha. HeLa cells were incubated in the presence or absence of 0.2 mM deferoxamine mesylate (DFO) for 4 hours at 37ºC. Nuclear extracts were prepared using the Nuclear/Cytosolic Fractionation Kit (#AKR-171). 100 pmol of non-biotinylated wild type or mutated HRE double stranded com-petitor oligos were added to the Complete DNA Binding Buffer just prior to inclusion in the assay.


HIF-1 Alpha DNA Binding Activity Assay Kit Colorimetric 96 Assays CBA-282

Cell hypoxia, or low oxygen condition, is a normal physiological response to certain body stressors such as high altitudes, but it also can be a symptom of pathological conditions. In some cases hypoxia may contribute to the inducement of oxidative stress. In response to hypoxic conditions, the hypoxia-inducible factor 1 transcriptional activator complex (HIF-1) plays a role in activating several hypoxia-responsive genes such as erythropoietin and VEGF. During hy-poxia, the alpha subunit of HIF-1 accumulates and translocates from the cytosol to the nucleus, where it dimerizes with the beta subunit and becomes tran-scriptionally active. It then binds transcriptional coacti-vators to induce gene expression. The HIF-1 Alpha DNA Binding Activity Assay Kit is an ELISA-based assay to detect activated HIF-1. Active HIF-1 complex is captured on a double-stranded oligo containing a hypoxic response element (HRE) that is attached to the plate. Detection is then performed with a primary antibody followed by an HRP-conjugated secondary antibody. The assay will detect HIF-1 complexes from human, mouse or rat samples.


HIF-1 Alpha Sandwich ELISA Kit Colorimetric 96 Assays CBA-280

HIF-1 Alpha Cell Based ELISA Kit Chemiluminescent 96 Assays CBA-281

HIF-1 Alpha ELISA Kits

Detection of Nuclear HIF-1 Alpha. HeLa cells were incubated in the presence or absence of 0.2 mM DFO for 4 hours at 37ºC. Nu-clear extracts were prepared using the Nuclear/Cytosolic Fractiona-tion Kit. HIF-1 Alpha levels were measured in untreated (blue bars) and treated (red bars) extracts according to the Assay Protocol.

Our HIF-1 Alpha ELISA Kits provide a convenient method for detection and quantitation of human, mouse, or rat HIF-1 Alpha in cells or tissues. Two ELISA kit formats are available: The HIF-1 Alpha Sandwich ELISA Kit detects HIF

-1 Alpha in any protein sample including tissue homogenates, whole cell lysates, or nuclear ex-tracts. Samples are added to an anti-HIF-1 Alpha antibody coated plate. Quantitation of unknown samples is performed by comparison of the OD values to those of a known standard.

The HIF-1 Alpha Cell Based ELISA Kit allows the detection of HIF-1 Alpha levels in intact cells. Cells are seeded in a tissue culture treated plate suitable for reading in a 96-well plate-based lumi-nometer. Cells are fixed and permeabilized to allow detection with the anti-HIF-1 antibody. De-tection is performed by chemiluminescence.

OXIDATIVE STRESS / DAMAGE ROS Assays

Reactive Oxygen Species Assays

Reactive oxygen species (ROS) such as superoxide and hydrogen peroxide are continually produced during metabolic processes. Excess ROS can lead to cellular injury in the form of damaged DNA, lipids and proteins. We offer assays for quantitation of various reactive oxy-gen species, in both in vitro and intracellular formats.

OxiSelect™ Intracellular ROS Assay Kit

The OxiSelect™ Intracellular ROS Assay Kit measures the activity of hydroxyl, peroxyl, and other reactive oxy-gen species. The assay uses the cell-permeable fluoro-genic probe DCFH-DA, which diffuses into cells and is deacetylated into the non-fluorescent DCFH. In the presence of ROS, the DCFH is oxidized into highly fluorescent DCF. Fluorescence is quantified on a fluorometric plate reader.

Sensitive: Detect concentrations as little as 10 pM Fast: Entire protocol takes about one hour


Fluorometric 96 Assays STA-342

5 x 96 Assays STA-342-5 OxiSelect™ Intracellular ROS Assay Kit

Assay Principle for the OxiSelect™ Intracellular ROS Assay.

Recent Product Citations 1. Koontz, J. et al. (2014). Competition through dimerization be-

tween antiapoptotic and proapoptotic HS-1-associated protein X-1 (Hax-1). J. Biol. Chem. 289:3468-3477.

2. Kim, E.Y. et al. (2013). NOX2 interacts with podocyte TRPC6 channels and contributes to their activation by diacylglycerol: essential role of podocin in formation of this complex. Am. J. Physiol. Cell Physiol. 305:C960-C971.

3. Abe, Y. et al. (2013). TGF-ß1 stimulates mitochondrial oxidative phosphorylation and generation of reactive oxygen species in cultured mouse podocytes, mediated in part by the mTOR path-way. Am. J. Phyisol. Renal Physiol. 305:F1477-F1490.

4. He, Q. et al. (2013). Tafazzin knockdown interrupts cell cycle progression in cultured neonatal ventricular fibroblasts. Am. J. Physiol. Heart Circ. Physiol. 305:H1332-H1343.

5. Kokkinaki, M. et al. (2013). Klotho regulates retinal pigment epithelial functions and protects against oxidative stress. J. Neu-rosci. 33: 16346-16359.

6. Hagan, C. et al. (2013). A common docking domain in progester-one receptor-B links DUSP6 and CK2 signaling to proliferative transcriptional programs in breast cancer cells. Nucleic Acids Res. 10.1093/nar/gkt706.

7. Teng, H. et al. (2013). Oxygen-sensitive mitochondrial accumu-lation of cystathione ß-synthase mediated by lon protease. PNAS 110:12679-12684.

8. Wang, H. et al. (2012). p53-induced gene 3 mediates cell death induced by glutathione peroxidase 3. J. Biol. Chem. 287:16890-16902.

9. Druz, A. et al. (2012). Glucose depletion activates MMU-mir-466h-5p expression through oxidative stress and inhibition of histone deacetylation. Nucleic Acids Res. 10.1093/nar/gks452.

10.Montalvo-Ortiz, B.L. et al. (2012). Characterization of EHOP-016, novel small molecule inhibitor of Rac GTPase. J. Biol. Chem. 287:13228-13238.






5 x 96 Assays STA-347-5 OxiSelect™ In Vitro ROS/RNS Assay Kit

OxiSelect™ In Vitro ROS/RNS Assay Kit

Free radicals and related reactive oxygen species (ROS) and reactive nitrogen species (RNS) can ap-pear in the body both inside and outside the cell. Until recently it has been difficult to detect ROS and RNS outside of intact cells. The OxiSelect™ In Vitro ROS/RNS Assay Kit allows you to measure ROS and RNS formation in various body fluids including urine, serum and plasma. It is also useful for testing cell lysates, tissue homoge-nates, and cell culture supernatants. The assay universally measures reactive oxygen and reactive nitrogen species that may include hydrogen peroxide, nitric oxide, peroxynitrite, peroxyl radicals, and others. The assay principle is similar to our Intra-cellular ROS Assay (previous page), except that the chemistry is modified to allow detection of ROS out-side the cell. Fluorescence is quantified on a fluoro-metric plate reader.

Sensitive: Detect concentrations as little as 10 pM for DCF or 40 nM for hydrogen peroxide

Fast: Entire protocol takes about one hour Versatile: Suitable for a wide variety of sample

types including urine, serum, plasma, cell lysates, tissue homogenates and cell culture supernatants

Recent Product Citations 1. Liu, X. et al. (2013). Epoxyeicosatrienoic acids prevent cisplatin-

induced renal apoptosis through a p38 mitogen-activated protein kinase-regulated mitochondrial pathway. Mol. Pharmacol. 84:925-934.


3. Xiao, D. et al. (2013). Estrogen normalizes perinatal nicotine-induced hypertensive responses in adult female rat offspring. Hypertension 61:1246-1252.

4. Wang, W. et al. (2012). Mono-(2-ethylhexyl) phthalate induces oxidative stress and inhibits growth of mouse ovarian antral follicles. Biol. Reprod. 87:152.

5. Ju, D. J. et al. (2012). Ethyl pyruvate ameliorates albuminuria and glomerular injury in the animal model of diabetic nephropa-thy. Am. J. Phyisol. Renal Physiol. 302:F606-F613.

6. Momi, S. et al. (2012). Nitric oxide enhances the anti-inflammatory and anti-atherogenic activity of atorvastatin in a mouse model of accelerated atherosclerosis. Cardiovasc. Res. 10.1093/cvr/cvs100.

7. Patterson, A.J. et al. (2011). Hypoxia-derived oxidative stress mediates epigenetic repression of PKC epsilon gene in foetal rat hearts. Cardiovasc. Res. 10.1093/cvr/cvr322.

8. Rathnasamy, G. et al. (2011). Iron and iron regulatory proteins in amoeboid microglial cells are linked to oligodendrocyte death in hypoxic neonatal rat periventricular white matter through produc-tion of proinflammatory cytokines and reactive oxygen/nitrogen species. J. Neurosci. 31:17982-17995.

Assay Principle for the OxiSelect™ In Vitro ROS/RNS Assay.



OxiSelect™ Hydrogen Peroxide Assay, Colorimetric

Hydrogen peroxide is one of the most prevalent and most stable of the various reactive oxygen species. The half-life of hydrogen peroxide is significantly longer than that of most ROS, making it easier to detect in many sample types.

Standard Curve Generated with the OxiSelect™ Hydrogen Peroxide/Peroxidase Assay (Fluorometric).


OxiSelect™ Hydrogen Peroxide Assay Kit Colorimetric 500 Assays STA-343


OxiSelect™ Hydrogen Peroxide/Peroxidase Assay Kit Fluorometric 500 Assays STA-344

*For the testing of hydrogen peroxide in cells and tissues, please see our OxiSelect™ Hydrogen Peroxide / Peroxidase Assay Kit below.

OxiSelect™ Hydrogen Peroxide / Peroxidase Assay, Fluorometric

Our OxiSelect™ Hydrogen Peroxide Assay Kit pro-vides a simple method for quantitation of hydrogen peroxide. This colorimetric assay measures the oxi-dation of ferrous (Fe2+) ions to ferric (Fe3+) ions in the presence of peroxides. The ferric ions form a com-plex with a provided dye which may be read on a standard microplate reader. The assay may be run with either aqueous phase or lipid phase samples.

Recent Product Citations 1. Kim, E.Y. et al. (2012). Sustained activation of N-methyl-D-

aspartate receptors in podocytes leads to oxidative stress, mobi-lization of transient receptor potential canonical 6 channels, nuclear factor of activated T cells activation, and apoptotic cell death. Mol. Pharmacol. 82:728-737.

2. Kim, E.Y. et al. (2012). Insulin increases surface expression of TRPC6 channels in podocytes: role of NADPH oxidases and reactive oxygen species. Am. J. Phyisol. Renal Physiol. 302:F298-F307.

Sensitive: Detect as little as 50 nM Fast: Easy 30 minute incubation Versatile: Measure either hydrogen peroxide or

peroxidase in plasma, serum, urine, cell culture supernatants, cell lysates and tissue homogenates

Our OxiSelect™ Hydrogen Peroxide / Peroxidase Assay Kit provides a convenient plate-based method for quantitation of hydrogen peroxide or peroxidases in a wide variety of sample types. This fluorometric assay uses a fluorogenic probe which is converted from a non-fluorescent to a fluo-rescent state in the presence of peroxides and is catalyzed by peroxidases. The kit includes both a hydrogen peroxide standard and a peroxidase standard for quantitative results with either target.

Sensitive: Detect as little as 1 µM Fast: Easy 30-90 minute incubation, depending on

sample type Versatile: Suitable for plasma, serum, urine, and

cell culture supernatants*



Direct detection: Probe binds directly to nitric ox-ide, not to by-products such as nitrate and nitrite

Sensitive: Detect as little as 3 nM Versatile: Read results as endpoint or time course

(kinetic) in a fluorescence plate reader, or visualize under a fluorescence microscope

OxiSelect™ Intracellular Nitric Oxide Assay Kit

OxiSelect™ In Vitro Nitric Oxide Assay Kits



5 x 96 Assays STA-800-5 OxiSelect™ Intracellular Nitric Oxide (NO) Assay Kit


OxiSelect™ In Vitro Nitric Oxide (Nitrite / Nitrate) Assay Kit



100 Assays STA-801

5 x 100 Assays STA-801-5 Fluorometric

Nitric oxide (NO) is a progenitor of various reactive nitrogen species (RNS) in conjunction with superox-ide anions via nitric oxide synthase (NOS). It plays a role in vascular diseases, diabetes, atherosclerosis, inflammatory diseases and cancer. Because of its short half-life, nitric oxide is often difficult to detect directly. The OxiSelect™ Intracellular Nitric Oxide Assay Kit allows direct detection of NO in intact cells. A cell-permeable fluorogenic probe is added to cells; upon treatment to induce oxidative stress, nitric oxide that is generated within the cell binds to the probe pro-ducing a bright fluorescent signal. Results may be visualized under a fluorescence microscope or quantified in a 96-well fluorescence plate reader. Induction of NOS in RAW 264.7 Cells. Cells were seeded in a

96-well plate at 100,000 cells/well. Cells were uninduced (left) or induced with 50 ng/mL LPS and 10 ng/mL IFN(right) for 20 hours at 37ºC.

Nitric oxide (NO) is difficult to detect directly in vitro due to its short half-life. It is therefore common to measure nitric oxide formation by detection of its final oxidized products, nitrate and nitrite. The OxiSelect™ In Vitro Nitric Oxide Assay Kits pro-vide a convenient plate-based method for the quanti-tation of nitrate and nitrite in a variety of sample types. First, nitrate is reduced to nitrite. Then total nitrite is measured by the addition of a Griess Re-agent (for colorimetric detection) or a fluorometric probe (for fluorescence detection). Results are then quantified in a 96-well plate reader. OxiSelect™ In Vitro Nitric Oxide Assay Kits are suit-able for use with serum, plasma, urine, saliva, cell lysates, and culture media.

Assay Principle for the OxiSelect™ In Vitro Nitric Oxide (Nitite / Nitrate) Assay, Fluorometric Format.

106

OXIDATIVE STRESS / DAMAGE Antioxidant Assays


Antioxidant Assays

ROS generation is normally counterbalanced by the action of antioxidant enzymes and other redox molecules. We offer two types of assays for antioxidant quantitation:

OxiSelect™ Catalase Activity Assay Kits



Fluorometric 96 Assays STA-339 OxiSelect™ Catalase Activity Assay Kit

Catalase is a ubiquitous enzyme that destroys hydro-gen peroxides formed during oxidative stress. Since hydrogen peroxides have a longer half-life than most free radicals and can make up a large portion of all reactive oxygen species, the ability to remove hydro-gen peroxides can be extremely important at combat-ing oxidative stress. Our OxiSelect™ Catalase Activity Assay Kits provide a quick, user-friendly protocol to monitor catalase activity from a variety of sample types. Kits are avail-able with either colorimetric or fluorometric detection.

Sensitive: Detect as little as 1.25 units/mL (colorimetric) or 50 mU/mL (fluorometric)

Fast: Obtain results in less than 30 minutes Versatile: Suitable for use with whole blood,

plasma, serum, cell lysates or tissue homogenates High Throughput: 96-well format

Assays to quantify the presence or activity of antioxidant molecules Assays to determine the antioxidant capacity of biomolecules

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

0 20 40 60 80 100

Catalase (U/mL)

OD

54

0n

m

Assay Principle for the OxiSelect™ Catalase Acitivity Assays. Catalase present in samples converts hydrogen peroxide into water and oxygen (Reaction 1). Any remaining hydrogen peroxide that is not converted reacts with a colorimetric or fluorometric probe in the presence of horseradish peroxidase (Reaction 2) to produce a color or fluorescence which is measured in a plate reader.

Standard Curve Generated with the OxiSelect™ Catalase Activity Assay Kit (Colorimetric).

Recent Product Citation Costantini, D. et al. (2013). Loss of integration is associated with reduced resistance to oxidative stress. J. Exp. Biol. 216:2213-2220. (STA-341)



OxiSelect™ Superoxide Dismutase Activity Assay

Superoxide dismutase (SOD), which catalyzes the dismutation of the superoxide anion into hydrogen peroxide and molecular oxygen, is one of the most important antioxidant enzymes.

Sensitive: Detect as little as 0.6 units/mL Fast: Obtain results in about 2 hours Versatile: Suitable for use with urine, serum, cells

or tissue samples


OxiSelect™ Superoxide Dismutase Activity Assay Colorimetric 100 Assays STA-340

Standard Curve Using the OxiSelect™ Superoxide Dismutase Activity Assay.

The OxiSelect™ Superoxide Dismutase Activity As-say uses a xanthine/xanthine oxidase (XOD) system to generate superoxide anions and a chromagen to produce a water-soluble dye upon reduction by the superoxide anions.

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

0.001 0.01 0.1 1 10 100

SOD (Units)

OD

490

nm

OxiSelect™ Superoxide Dismutase Activity Assay Principle. Superoxide anions generated by a Xanthine/Xanthine Oxidase system are detected with the provided chromagen. SOD reduces superoxide concentrations, so higher SOD concentrations result in a decreased signal.

Recent Product Citations 1. Wysocki, J. et al. (2014). ACE2 deficiency increases NADPH-

mediated oxidative stress in teh kidney. PHY2 2:e00264. 2. Yin, J. et al. (2014). Development of an antioxidant system after

early weaning in piglets. J. Anim. Sci. 92:612-619. 3. Gong, E.J. et al. (2013). Low-dose-rate radiation exposure leads

to testicular damage with decreases in DNMT1 and HDAC1 in the murine testis. J. Radiat. Res. 10.1093/jrr/rrt090.


5. Paneni, F. et al. (2013). Deletion of the activated protein-1 tran-scription factor JunD induces oxidative stress and accelerates age-related endothelial dysfunction. Circulation 127:1229-1240.

6. Connell, B.J. et al. (2012). UPEI-100, a conjugate of lipoic acid and apocynin, mediates neuroprotection in a rat model of ische-mia/reperfusion. Am. J. Physiol. Regulatory Integrative Comp. Physiol. 302:R866-R895.

7. Zhang, Z. et al. (2012). TRPM2 Ca2+ channel regulates energy balance and glucose metabolism. Am. J. Phyisol. Endocrin. Metab. 302:E807-E816.



OxiSelect™ Total Glutathione (GSSG/GSH) Assay Kit

Glutathione is an intracellular tripeptide thiol that pro-tects cells from free radicals by acting as an antioxi-dant. Glutathione exists within cells in both reduced (GSH) and oxidized (GSSG) forms; it is involved in the breakdown of peroxides and also helps maintain exogenous antioxidants such as vitamins C and E. The OxiSelect™ Total Glutathione Assay Kit is a quantitative assay for measuring total combined GSH and GSSG content in a variety of sample types. Oxi-dized glutathione is enzymatically reduced, followed by colorimetric detection in a microplate reader.

Sensitive: Detect as little as 8 nM total glutathione Fast: Obtain results in less than 30 minutes Versatile: Suitable for use with saliva, urine,

serum, plasma, and cell or tissue lysates


OxiSelect™ Total Glutathione (GSSG/GSH) Assay Kit Colorimetric 100 Assays STA-312

OxiSelect™ Glutathione Reductase Assay Kit


OxiSelect™ Glutathione Reductase Assay Kit Colorimetric 100 Assays STA-812

Assay Principle for the OxiSelect™ Total Glutathione Assay Kit. In the presence of NADPH, glutathione reductase (provided) con-verts all glutathione into reduced form (GSH). The reduced glutathione then reacts with the provided chromogen to yield a color detect-able at 405 nm.

Standard Curve Generated with the OxiSelect™ Glutathione Reductase Assay Kit. Various concentrations of glutathione reductase standard were tested according to the Assay Protocol. OD values were read at 1 minute increments at 405 nm.

The OxiSelect™ Glutathione Reductase Assay Kit is a quantitative assay for measuring the activity levels of glutathione reductase in a variety of sample types. The assay principle is similar to that of our Total Glu-tathione Assay Kit above, except that endogenous levels of glutathione reductase drive the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH).

Sensitive: Detect activity levels as low as 0.6 mU/mL

Fast: Obtain results in less than 30 minutes Versatile: Suitable for use with erythrocytes,

plasma, cell lysates, or tissue extracts

Recent Product Citations 1. Mani, S. et al. (2013). Decreased endogenous production of

hydrogen sulfide accclerates atherosclerosis. Circulation 127:2523-2534.

2. Karakus, E. et al. (2013). Agomelatine: an antidepressant with new potent hepatoprotective effects on paracetamol-induced liver damage in rats. Human and Exp. Toxicol. 10.01177/0960327112472994.



OxiSelect™ Cellular Antioxidant Assay Kit, for in vivo Evaluation of Exogenous Antioxidants


OxiSelect™ Cellular Antioxidant Activity Assay Kit Fluorometric 192 Assays STA-349

Measuring the effects of antioxidant compounds in an in vitro assay may not accurately reflect their efficacy because such assays do not account for physiologi-cal conditions such as pH, temperature, uptake, me-tabolism, or the bioavailability or efficacy of an anti-oxidant compound. The OxiSelect™ Cellular Antioxidant Activity Assay Kit provides a mechanism to test exogenous antioxi-dants in a cell-based environment, delivering a more accurate measurement of the compound’s true physiological efficacy. A cell-permeable fluorometric dye is added to intact cells; when free radicals are generated, they bind to the dye producing a bright fluorescent signal. When the exogenous antioxidant is added, it eliminates the free radicals resulting in decreased fluorescence.

Physiological: Measures the efficacy of an antioxi-dant compound in a cellular environment

More Accurate: Compared to in vitro assays that do not take into account pH, temperature, or other relevant intracellular conditions

Sensitive: Detects as little as 10 µM Quercetin, a bioflavonoid which serves as the assay’s standard

Fast: Obtain results in about one hour

0

2

4

6

8

10

12

14

16

18

0 10 20 30 40 50 60 70

Flu

ore

scen

ce (

RF

Us)

Time (Minutes)

2000 uM Quercetin

1000

500

250

125

62.5

31.3

0

Cellular Antioxidant Activity of Quercetin in HeLa Cells. 60,000 HeLa cells were seeded and cultured in a 96-well plate until confluent. Cells were then pretreated with DCFH-DA and Quercetin for 60 minutes at 37ºC. Free Radical Initiator was then added to the cells to begin the assay. Fluorescence readings were taken every 5 minutes for one hour at 37ºC.

Assay Principle for the OxiSelect™ Cellular Antioxidant Activ-ity Assay Kit. An exogenous antioxidant compound is added to cells along with DCFH-DA dye. Upon entry into the cell, the DCFH-DA is cleaved to DCFH which can bind reactive oxygen species (ROS) generated within the cell by the addition of a free radical initiator. Binding of DCFH to ROS yields DCF which produces a bright fluorescence. The presence of the exogenous antioxidant compound reduces the ROS available to the DCFH dye, yielding a lower fluorescent signal.



OxiSelect™ ORAC and HORAC Activity Assay Kits

The ORAC (Oxygen Radical Antioxidant Capacity) and HORAC (Hydroxyl Radical Antioxidant Capac-ity) assays measure the antioxidant capacity of bio-molecules against peroxyl radicals and hydroxyl radicals, respectively. The assays are suitable for plasma, cell fractions, and tissue lysates, as well as solid and aqueous nutrition samples.

Assay Principle for the OxiSelect™ Oxygen Radical Antioxidant Capacity (ORAC) Assay.




OxiSelect™ HORAC Activity Assay Kit Fluorometric 192 Assays STA-346


OxiSelect™ ORAC Activity Assay Kit

Recent Product Citations 1. Ungvari, Z. et al. (2013). Testing predictions of the oxidative

stress hypothesis of aging using a novel invertebrate model of longevity: the giant clam (Tridacna derasa). J. Gerontol. A. Biol. Sci. Med. Sci. 68:359-367.

2. Bailey-Downs, L.C. et al. (2011). Liver-specific knockdown of IGF-1 decreases vascular oxidative stress resistance by impair-ing the Nrf2-dependent antioxidant response: a novel model of vascular aging. J. Gerontol. A. Biol. Sci. Med. Sci. 10.1093/gerona/glr164.


OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit Colorimetric 200 Assays STA-360

Recent Product Citations 1. Stark, M. et al. (2013). Differential effects of docosahexaenoic

acid on preterm and term placental pro-oxidant/antioxidant bal-ance. Reproduction 146:243-251.

2. Bakalova, R. et al. (2013). Tissue redox activity as a hallmark of carcinogenesis: from early to terminal stages of cancer. Clin. Cancer Res. 19:2503-2517.

3. Wang, Y. et al. (2013). Therapeutic effect of MG-132 on diabetic cardiomyopathy is associated with its suppression of protea-somal activities: roles of Nrf2 and NF-kB. Am. J. Physiol. Heart Circ. Physiol. 304:H567-H578.

OxiSelect™ Total Antioxidant Capacity Assay Principle.

OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit

The OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit measures the total antioxidant capacity of biomolecules from a variety of sample types via a Single Electron Transfer (SET) mechanism. The as-say works with a variety of antioxidants and is suit-able for testing plasma, serum, urine, cell lysates, tissue homogenates and food extracts.

112

CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein

Small GTPase Activation Assay Principle.

Safe: Non-radioactive assay format Visual Check: Agarose beads can be easily seen Fast Results: 1 hour plus electrophoresis/blotting

Small GTP-binding proteins (GTPases) regulate a variety of cell signaling pathways and are therefore involved in a wide range of cell functions, processes, and morphology. The most studied small GTPases include Ras, Rac, Rho and Cdc42. We offer a variety of tools to enable the study of these small GTPase family members:

In addition, we offer sensitive assays to detect cyclic AMP and cyclic GMP, both of which are important regulators in the G-Protein signaling cascade.

Small GTPase / G-Protein Signaling

Small GTPase Activation Assays Small GTPase Activation ELISA Kits Small GTPase Assay Beads Active Rac-GEF Assay

Small GTPase Expression Vectors Small GTPase Premade Adenoviruses Small GTPase Retroviral Constructs Small GTPase Recombinant Proteins


Small GTPase Activation Assays

Our Small GTPase Activation Assays use visible aga-rose beads to selectively pull down the active form of the target of interest. The precipitated GTPase is then detected by Western blot using a target specific anti-body included in the kit. If you are studying more than one small GTPase tar-get, consider one of our Small GTPase Activation Assay Combo Kits. These combo kits allow you to measure the following targets at a savings compared to buying separate kits for each target: Rac1 and Cdc42 RhoA, Rac1 and Cdc42

Visible Agarose Beads Provided in the Small GTPase Activation Assays. Beads are easy to visual-ize, making it easier to avoid poten-tial loss during washes and aspira-tions.

113

Small GTPase/G-Protein

Small GTPase Activation Assays (continued)


Cdc42 Activation Assay Immunoblot/ECL 20 Assays STA-402

Rac1 Activation Assay Immunoblot/ECL 20 Assays STA-401-1

Ral Activation Assay Immunoblot/ECL 20 Assays STA-408

Ran Activation Assay Immunoblot/ECL 20 Assays STA-409

Pan-Ras Activation Assay Immunoblot/ECL 20 Assays STA-400

RhoA Activation Assay Immunoblot/ECL 20 Assays STA-403-A

Rac1/Cdc42 Activation Assay Combo Kit Immunoblot/ECL 20 Assays/Target STA-404

RhoA/Rac1/Cdc42 Activation Assay Combo Kit Immunoblot/ECL 10 Assays/Target STA-405

Rac2 Activation Assay Immunoblot/ECL 20 Assays STA-401-2

RhoC Activation Assay Immunoblot/ECL 20 Assays STA-403-C

Arf6 Activation Assay Immunoblot/ECL 20 Assays STA-407-6

Arf1 Activation Assay Immunoblot/ECL 20 Assays STA-407-1

Rap1 Activation Assay Immunoblot/ECL 20 Assays STA-406-1

RhoB Activation Assay Immunoblot/ECL 20 Assays STA-403-B

Rap2 Activation Assay Immunoblot/ECL 20 Assays STA-406-2

H-Ras Activation Assay Immunoblot/ECL 20 Assays STA-400-H

K-Ras Activation Assay Immunoblot/ECL 20 Assays STA-400-K

N-Ras Activation Assay Immunoblot/ECL 20 Assays STA-400-N

Recent Product Citations 1. Ohashi, K. et al. (2012). Lung cancers with acquired resistance to EGFR inhibitors occasionally harbor BRAF gene mutations but lack mu-

tations in KRAS, NRAS or MEK1. PNAS 109:E2127-E2133. (STA-400) 2. Geryk-Hall, M. et al. (2010). Driven to death: inhibition of farnesylation increases Ras activity in osteosarcoma and promotes growth arrest

and cell death. Mol. Cancer Ther. 9:1111-1119. (STA-400) 3. Camalier, C.E. et al. (2010). Elevated phosphate activates N-ras and promotes cell transformation and skin tumorigenesis. Cancer Prev.

Res. 3:359-370. (STA-400) 4. Tsukamoto, Y. et al. (2013). A novel heart failure mice model of hypertensive heart disease by angiotensin II infusion, nephrectomy, and

salt loading. Am. J. Physiol. Heart Circ. Physiol. 305:H1658-H1667. (STA-401-1) 5. Petzold, T. et al. (2013). ß1 integrin-mediated signals are required for platelet granule secretion and hemostasis in mouse. Blood 122:2723-

2731. (STA-401-1) 6. Cristante, E. et al. (2013). Identification of an essential endogenous regulator of blood-brain barrier integrity, and its pathological and thera-

peutic implications. PNAS 110:832-841. (STA-401-1) 7. Yanagashita, T. et al. (2014). Actin-binding protein, espin: a novel metastatic regulator for melanoma. Mol. Cancer Res. 12:440-446. (STA-

401-1, STA-403-A) 8. He, S. et al. (2013). SRGAP1 is a candidate gene for papillary thyroid carcinoma susceptibility. J. Clin. Endocrinol. Metab. 98:E973-E980.

(STA-402) 9. Fiuza, M. et al. (2013). GluN3A expression restricts spine maturation via inhibition of GIT1/Rac1 signaling. PNAS 110:20807-20812. (STA-

403-A) 10. Basu, M. et al. (2012). Wnt/ß-catenin pathway is regulated by PITX2 homeodomain protein and thus contributes to the proliferation of hu-

man ovarian adenocarcinoma cell SKOV-3. J. Biol. Chem. 288:4355-4367. (STA-403-A) 11. Holmes, K.M. et al. (2012). Insulin-like growth factor-binding protein 2-driven glioma progression is prevented by blocking a clinically signifi-

cant integrin, integrin-linked kinase, and NF-kB network. PNAS 109:2168-2173. (STA-404) 12. Wang, J. et al. (2013). DEK depletion negatively regulates Rho/ROCK/MLC pathway in non-small cell lung cancer. J. Histochem. & Cyto-

chem. 10.1369/00221155413488120. (STA-405) 13. Quint, P. et al. (2013). Sphingosine 1-phosphate (S1P) receptors 1 and 2 coordinately induce mesenchymal cell migration through S1P

activation of complementary kinase pathways. J. Biol. Chem. 288:5398-5406. (STA-405) 14. Baranwai, S. et al. (2011). Molecular characterization of the tumor-suppressive function of nischarin in breast cancer. J. Natl. Cancer Inst.

10.1093/jnci/djr350. (STA-405) 15. Grossman, A. et al. (2013). The small GTPase ARF6 stimulates ß-catenin transcriptional activity during WNT5A-mediated melanoma inva-

sion and metastasis. Sci Signal 6:ra14. (STA-407-6)


CELL SIGNALING & PROTEIN BIOLOGY

114


Active Rac-GEF Assay Kit (Tiam1) Immunoblot/ECL 20 Assays STA-422

Active Rac-GEF Assay Kit (Tiam1)

Guanine nucleotide exchange factors (GEFs) acti-vate small GTPases by catalyzing the exchange of GDP for GTP. Our Active Rac-GEF Assay Kit (Tiam1) uses the agarose bead technology of our Small GTPase Activation Assays (previous page). Agarose beads pull down the active form of Rac-GEFs from en-dogenous lysates or purified samples. The specific GEF known as Tiam1 is then specifically detected with a polyclonal antibody.



Recent Product Citation Oubaha, M. et al. (2012). Formation of a PKC/ß-catenin complex in endothelial cells promotes angiopoietin-1-induced collective direc-tional migration and angiogenic sprouting. Blood 120:3371-3381.

Tiam1 Activation Assay. Left: 293 cells were transfected with active Tiam1. Active Tiam1 in lysate was pulled down with Rac1 G15A agarose beads. Right: Active Tiam-1 in 2 mg of MDA-231 cell lysate was pulled down with Rac1 G15A agarose beads and probed with anti-Tiam1 anti-body.

96-Well Ras Activation ELISA Kits



Chemiluminescent 96 Assays STA-441 96-Well Ras Activation ELISA Kit

Our 96-Well Ras Activation Assays use the Raf1 Rho binding domain (Raf1 RBD) to selectively pull down the active form of Ras from purified or endogenous samples. The captured GTP-Ras is then detected by a pan-Ras antibody and HRP-conjugated secondary antibody. Detection is by either colorimetric or chemiluminescent plate reader.

EGF Stimulation and Active Ras Detection with the 96-Well Ras Activation ELISA Kit. HeLa cells were serum starved for 18 hours before EGF stimulation of 50 ng/mL for 2 minutes. Lysates were then prepared according to the assay protocol.

Pan-Ras Antibody Specificity. Anti-pan-Ras antibody reactivity with H-Ras, K-Ras and N-Ras human isoforms by dot blot.

115

Product Name Target Size Catalog Number

Cdc42 G15A Agarose Beads Cdc42-GEF 800 µg STA-433

Rac1 G15A Agarose Beads Rac1-GEF 800 µg STA-432

RhoA G17A Agarose Beads RhoA-GEF 400 µg STA-431

GEF Agarose Assay Beads

Recent Product Citations 1. Yuen, H. et al. (2013). RanGTPase: a candidate for Myc-

mediated cancer progression. J. Natl. Cancer Inst. 105:475-488. (STA-410)

2. Moniz, S. et al. (2007). Protein kinase WNK2 inhibits cell prolif-eration by negatively modulating the activation of MEK1/ERK1/2. Oncogene 26(41):6071-6081. (STA-410)

3. Pothula, S. et al. (2013). Regulation of Cdc42 expression and signaling is critical for promoting corneal epithelial wound heal-ing. Invest. Ophthalmol. Vis. Sci. 54:5343-5352. (STA-411)

4. Zhang, Q-G. et al. (2009). Estrogen attenuates ischemic oxida-tive damage via an estrogen receptor alpha-mediated inhibition of NADPH oxidase activation. J. Neurosci. 29:13823-13836. (STA-411)

5. Levy-Adam, F. et al. (2008). Heparanase facilitates cell adhe-sion and spreading by clustering of cell surface heparan sulfate proteoglycans. PLoS ONE 3(6):e2319. (STA-411, STA-412)

6. Sabbatini, M. E. et al. (2010). CCK activates RhoA and Rac1 differentially through G-alpha-13 and G-alpha-q in mouse pan-creatic acini. Am. J. Physiol. Cell Physiol. 298:C592-C605. (STA-411, STA-412)

7. Sabbatini, M. et al. (2008). Rap1 activation plays a regulatory role in pancreatic amylase secretion. J. Biol. Chem. 283:23884-23894. (STA-412)


Small GTPase/G-Protein CELL SIGNALING & PROTEIN BIOLOGY

Small GTPase Agarose Assay Beads

Our agarose beads are useful for selectively pulling down only the active form of small GTPases. The beads are colored for easily visualization. These are the same beads used in our Small GTPase Ac-tivation Assays (p. 108-109).

Visible Agarose Beads. Beads are easy to visualize, making it easier to avoid potential loss during washes and aspirations.

Recent Product Citations 1. Ngok, S. et al. (2013). Phosphorylation-mediated 14-3-3 protein

binding regulates the function of the Rho-specific guanine nu-cleotide exchange factor (RhoGEF) Syx. J. Biol. Chem. 288:6640-6650. (STA-431)

2. Colacios, C. et al. (2011). The p.Arg63Trp polymorphism con-trols Vav1 functions and Fox3p regulatory T cell development. J. Exp. Med. 208:2183-2191. (STA-432)

Product Name Target Size Catalog Number

GGA3 PBD Agarose Beads Arf 400 µg STA-419

PAK1 PBD Agarose Beads Cdc42, Rac 400 µg STA-411

Raf1 RBD Agarose Beads Ras 400 µg STA-410

RalBP1 PBD Agarose Beads Ral 400 µg STA-420

RalGDS RBD Agarose Beads Rap 400 µg STA-418

RanBP1 Agarose Beads Ran 400 µg STA-421

Rhotekin RBD Agarose Beads Rho 400 µg STA-412

Our GEF agarose beads are useful for selectively pulling down only the active form of guanine nu-cleotide exchange factors (GEF). The beads are colored for easily visualization.



Product Name Vectors Size Catalog Number

Cdc42 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 100 µL STA-455

GFP-Cdc42 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 100 µL STA-451

Rac1 Expression Vector Set Wild Type, T17N (Dom. Neg.), G12V (Active) 3 x 100 µL STA-454

GFP-Rac1 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 100 µL STA-450

H-Ras Expression Vector Set Wild Type, T17N (Dom. Neg.), G12V (Active) 3 x 100 µL STA-457

RhoA Expression Vector Set Wild Type, T19N (Dom. Neg.), G14V (Active) 3 x 100 µL STA-456

GFP-RhoA Expression Vector Set Wild Type, T19N (Dom. Neg.), Q63L (Active) 3 x 100 µL STA-452


3 x 100 µL STA-460 Exoenzyme C3 Expression Vector

Small GTPase Expression Vector Sets

Our Small GTPase Expression Vectors are ideal tools for the study of the most commonly researched small GTPase targets. Each set contains 3 mammal-ian expression vectors: Wild type Dominant negative Constitutively active Vectors are available with or without a GFP reporter gene. All vectors are supplied as bacterial glycerol stocks.

Exoenzyme C3 (Rho Inhibitor) Expression Vector

Product Name Vectors Size Catalog Number

Active Rac1 Expression Vector Set Q61L, Q61L/F37A, Q61L/Y40C 3 x 100 µL STA-458

Active H-Ras Expression Vector Set V12, V12/S35, V12/C40 3 x 100 µL STA-459

Active Small GTPase Expression Vector Sets

Our Active Small GTPase Expression Vectors are similar to the expression vectors above, except that they are provided as sets of 3 different active mutants for a single small GTPase target. All vectors are supplied as bacterial glycerol stocks.

This vector is supplied as bacterial glycerol stock.


Recent Product Citation Zhao, B. et al. (2012). TNF-induced osteoclastogenesis and inflam-matory bone resorption are inhibited by transcription factor RBP-J. J. Exp. Med. 209:319-334. (RTV-101)


Small GTPase Recombinant Adenoviruses


Cdc42 ADV-152



Rac1 ADV-149





Ras V12C40 ADV-148

Ras V12S35 ADV-147



All of Cell Biolabs’ premade recombinant adenovi-ruses contain 5 x 109 viral particles per vial. They are provided as 50 µL aliquots at a concentration of 1 x 1011 viral particles/mL in TBS with 10% glycerol.

Actin Cytoskeleton Staining. Cos-7 cells were infected with purified Ad-Ras V12 (ADV-146) at 50 MOI (multiplicity of infection). Membrane ruffling was visual-ized by staining the actin cytoskeleton with Rhodamine-coupled Phalloidin.

Recent Product Citations 1. Black, S.A. et al. (2008). TGFß1 stimulates connective tissue

growth factor (CCN2/CTGF) expression in human gingival fibroblasts through a RhoA-independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGFß1-induced CCN2/CTGF expression. J. Biol. Chem. 283:10835-10847. (ADV-145, ADV-150, ADV-153, ADV-156)

2. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires avß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Biol. Cell 23:1104-1114. (ADV-150)

3. Thomas, M.A. et al. (2009). E4orf1 limits the oncolytic potential of the E1B-55K-deleted adenovirus. J. Virol. 83:2406-2416. (ADV-150)

4. Rendon, B. et al. (2007). Regulation of human lung adenocarci-noma cell migration and invasion by MIF. J. Biol. Chem. 282:29910-29918. (ADV-150)

5. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150, ADV-153, ADV-154)

6. Cheng, Z.-J. et al. (2010). Co-regulation of caveolar and Cdc42-dependent fluid phase endocytosis by phosphocaveolin-1. J. Biol. Chem. 285:15119-15125. (ADV-153)

7. Perez-Moreno, M. et al. (2008). Loss of p120 catenin and links to mitotic alterations, inflammation, and skin cancer. PNAS 105:15399-15404. (ADV-156)

8. Neal, M. et al. (2013). A critical role for TLR4 induction of auto-phagy in the regulation of enterocyte migration and the patho-genesis of necrotizing enterocolitis. J. Immunol. 190:3541-3551. (ADV-156, ADV-157)

9. Vaught, D. et al. (2009). Regulation of mammary gland branch-ing morphogenesis by EphA2 receptor tyrosine kinase. Mol. Biol. Cell 20:2572-2581. (ADV-157)

10.Brantley-Sieders, D. et al. (2008). The receptor tyrosine kinase EphA2 promotes mammary adenocarcinoma tumorigenesis and metastatic progression in mice by amplifying ErbB2 signal. J. Clin. Invest. 118:64-78. (ADV-157)


N-Ras K61 pBABEpuro RTV-222

Ras V12 pBABEpuro RTV-101

Ras V12C40 pBABEpuro RTV-104

Ras V12G37 pBABEpuro RTV-103

Ras V12S35 pBABEpuro RTV-102

RhoA L63 pBABEhygro RTV-204

Our recombinant retroviral plasmids contain a spe-cific gene cloned into a pBABE vector backbone. Each vector is supplied as 100 µL of bacterial glycerol stock.

Gene-Specific Recombinant Retroviral Vectors


Cdc42 L61 pBABEhygro RTV-203

myr-Rac1 pBABEpuro RTV-201

Rac1 V12 pBABEhygro RTV-202

Rac3 V12 pBABEhygro RTV-205

K-Ras pBABEpuro RTV-220

K-Ras Q61 pWZLhygro RTV-221



Small GTPase Recombinant Human Proteins

Regulators of G Protein Signaling (RGS) Recombinant Human Proteins

Protein Name Tag / Location Catalog Number

Cdc42 T7 / N-term STA-700

Rab1a 6xHis / N-term STA-701

Rab1b 6xHis / N-term STA-702



Rab3b 6xHis / N-term STA-705

Size

50 µg

10 µg

10 µg

20 µg

20 µg

20 µg

Rab3d 6xHis / N-term 20 µg STA-706

Rab4a 6xHis / N-term 10 µg STA-707

Rab5a None 20 µg STA-708

Rab5b 6xHis / N-term 20 µg STA-709





Rab13 6xHis / N-term 10 µg STA-714



Rab18 T7 / N-term 10 µg STA-717









Protein Name Tag / Location Size Catalog Number

Rab 35 6xHis / N-term 10 µg STA-726

Rab IF 6xHis / N-term 10 µg STA-727

Rac1 6xHis / N-term 50 µg STA-728

Rac1 None 50 µg STA-729

Rac2 6xHis / N-term 20 µg STA-730

Rac3 None 20 µg STA-731

Ral A 6xHis / N-term 25 µg STA-732

Ral B 6xHis / N-term 10 µg STA-733

Ran 6xHis / N-term 10 µg STA-734

Rap1a 6xHis / N-term 10 µg STA-735

Rap1b 6xHis / N-term 10 µg STA-736

Rap2a 6xHis / N-term 10 µg STA-737

RERG 6xHis / N-term 10 µg STA-738

RHEB T7 / N-term 50 µg STA-739

RhoA 6xHis / N-term 20 µg STA-740

RhoB 6xHis / N-term 10 µg STA-741

RhoC 6xHis / N-term 10 µg STA-742

RhoD 6xHis / N-term 5 µg STA-743

RND1 6xHis / N-term 20 µg STA-744

RND3 6xHis / N-term 20 µg STA-745

SAR1A 6xHis / N-term 20 µg STA-746

H-Ras 6xHis / C-term 25 µg STA-747

K-Ras 6xHis / N-term 25 µg STA-748

N-Ras None 10 µg STA-749

R-Ras 6xHis / N-term 10 µg STA-750

R-Ras2 6xHis / N-term 50 µg STA-751


RGS2 6xHis / N-term 5 µg STA-780










Recombinant GRP-PH Domain


100 µg STA-200

1 mg STA-200-1MG Recombinant GRP-PH Domain

The GRP1 (general receptor for phosphoinositide) protein binds phosphatidylinositol-3,4,5-triphosphate (PIP3) through a pleckstrin homology (PH) domain and displays a region of high sequence similarity to the yeast Sec7 protein. This recombinant protein is expressed and purified from E. coli as a fusion protein, and is provided at 1.0 mg/ml in 1X PBS.

Cyclic AMP and GMP ELISA Kits

Sensitive: Detect as little as 1 pmol/mL Versatile: Suitable for use with cell and tissue lys-

ates, urine, plasma, or culture medium Convenient: Strip-well plate format with either

colorimetric or chemiluminescent detection

Cyclic AMP and cyclic GMP are important regulatory molecules in the GPCR signaling cascade. Our cAMP and cGMP ELISA Kits provide a highly sensitive method to measure low levels of cAMP or cGMP in a variety of sample types.


cAMP ELISA Kit



Chemiluminescent 96 Assays STA-501


cGMP ELISA Kit



96 Assays STA-506

5 x 96 Assays STA-506-5 Chemiluminescent

0

0.5

1

1.5

2

2.5

0.0 0.1 1.0 10.0 100.0 1000.0 10000.0 100000.0

cAMP (pmol/mL)

OD

45

0n

m

Standard Curve Created with the cAMP ELISA Kit, Colorimetric Format.

Recent Product Citations 1. Meyer, R. et al. (2013). GPR37 and GPR37L1 are receptors for

the neuroprotective and glioprotective factors prosaptide and prosaposin. PNAS 110:9529-9534. (STA-500)

2. Sun, Z. et al. (2011). The WD40 repeat protein WDR26 binds Gßg and promotes Gßg-dependent signal transduction and leukocyte migration. J. Biol. Chem. 286:43902-43912. (STA-500)

3. Chen, M. et al. (2010). Involvement of cAMP in nerve growth factor-triggered p35/Cdk5 activation and differentiation in PC12 cells. Am. J. Physiol. Cell Physiol. 299:C516-C527. (STA-500)

4. McCoy, K.L. et al. (2010). PAR1 and PAR2 couple to overlap-ping and distinct sets of G proteins and linked signaling path-ways to differentially regulate cell physiology. Mol. Pharmacol. 77:1005-1015. (STA-500)

5. Liu, L. et al. (2012). Radil controls neutrophil adhesion and motil-ity through ß2-integrin activation. Mol. Biol. Cell 23:4751-4765. (STA-501)



CELL SIGNALING & PROTEIN BIOLOGY Kinase Assays

Rho Kinase (ROCK) Activity Assays

Rho Kinase (ROCK) is a serine/threonine kinase which is a target of Rho. ROCK mediates Rho signaling and reorganizes the actin cytoskeleton via the phosphorylation of several substrates that contribute to contractility and the assembly of actin filaments.


ROCK Activity Immunoblot Kit Immunoblot 20 Assays STA-415

96-Well ROCK Activity Assay 96 Assays STA-416


Results Using the ROCK Activity Immunoblot Kit. Lanes 1, 3, 5, 7: Without ROCK (negative control). Lanes 2, 4, 6, 8: With ROCK. Lanes 1 & 2: 200 ng MYPT1; Lanes 3 & 4: 100 ng; Lanes 5 & 6: 50 ng; Lanes 7 & 8: 25 ng. Phosphorylation of MYPT1 substrate was detected by anti-phospho-MYPT1 as de-scribed in the protocol.

96-Well ROCK Activity Assay Principle.

Our ROCK Activity Assays provide a non-radioactive format to measure the level of active ROCK in cell or tissue lysates. The immunoblot kit provides a conven-ient format for measuring ROCK activity in a few sam-ples, while the 96-well Activity Assay contains a strip-well plate precoated with MYPT1 for higher throughput.

Recent Product Citations 1. Georgess, D. et al. (2014). Comparative transcriptomics reveals

RhoE as a novel regulator of actin dynamics in bone-resorbing osteoclasts. Mol. Biol. 25:380-396. (STA-415)

2. Wang, J.N. et al. (2011). Response gene to complement 32 pro-motes vascular lesion formation through stimulation of smooth muscle cell proliferation and migration. Arterioscler. Thromb. Vasc. Biol. 31:e19-e26. (STA-415)

3. Xiao, L. et al. (2009). ROCK mediates phorbol ester-induced apoptosis in prostate cancer cells via p21-Cip1 upregulation and JNK. J. Biol. Chem. 284:29365-29375. (STA-415)

4. Burger, D. et al. (2011). Endothelial microparticle formation by angiotensin II is mediated via ang II receptor type I/NADPH oxi-dase/Rho kinase pathways targeted to lipid rafts. Arterioscler. Thromb. Vasc. Biol. 31:1898-1907. (STA-416)

5. Haas, B. et al. (2009). Protein kinase G controls brown fat cell differentiation and mitochondrial biogenesis. Sci. Signal. 2:ra78. (STA-416)


Kinase Assays CELL SIGNALING & PROTEIN BIOLOGY

Checkpoint Kinase Activity Assays

Checkpoint kinases, including CHK1 and CHK2, can be activated in response to DNA damage prior to mi-tosis. These kinases phosphorylate Cdc25C, a pro-tein phosphatase, at Ser-216. This phosphorylation ultimately leads to cell cycle arrest, preventing mitosis and avoiding the passage of DNA damage to daugh-ter cells. Our Checkpoint Kinase Activity Assays allow you to conveniently measure the activity of CHK1 and CHK2. The assays use recombinant Cdc25C as a checkpoint kinase substrate. Phosphorylated Cdc25C (Ser216) is detected using a phospho-specific antibody. Checkpoint Kinase Activitiy As-says are available in two formats: a Western blot assay and a 96-well plate-based activity assay.


Checkpoint Kinase Activity Immunoblot Kit Immunoblot 20 Assays STA-413

96-Well Checkpoint Kinase Activity Assay Kit 96 Assays STA-414


96-Well Checkpoint Kinase Activity Assay Principle.

CHK1 Activity Using the Checkpoint Kinase Activity Immunoblot Kit. Lane 1: Negative Control; Lane 2: 10 ng of active CHK1.

122

GFP Quantitation Kit


GFP Quantitation Kit Fluorometric 100 Assays AKR-120

Most imaging studies of rGFP are qualitative, and quantitation by FACS is time consuming and expen-sive. Our GFP ELISA kit uses a standard microplate reader to quantify GFP levels with extremely high sensitivity. This kit will detect GFP form Aequorea victoria as well as its variants.

Standard Curve Generated with the GFP ELISA Kit.

Sensitive: Detection limit of 30 pg/mL Versatile: Quantify GFP and its variants: BFP,

CFP or YFP Easy Quantitation: Measure GFP levels in a

standard microplate reader

Recent Product Citations 1. Mango, R. et al. (2014). C-C chemokine receptor 5 on pulmo-

nary mesenchymal cells promotes experimental metastasis via the induction of erythroid differentiation regulator 1. Mol. Cancer Res. 12:274-282.

2. Mitchell, A. et al. (2014). Promyelocytic leukemia protein is a cell-intrinsic factor inhibiting parvovirus DNA replication. J. Virol. 88:925-936.

3. Coghill, J.M. et al. (2013). CC chemokine receptor 8 potentiates donor Treg survival and is critical for the prevention of murine graft-versus-host disease. Blood 122:825-836.

4. Pedersen, J. et al. (2012). Glucose metabolism is altered after loss of L cells and -cells but not influenced by loss of K cells. Am. J. Physiol. Endocrinol. Metab. 304:E60-E73.

5. Zhou, W. et al. (2012). Inducible podocyte injury and proteinuria in transgenic Zebrafish. J. Am. Soc. Nephrol. 23:1039-1047.

6. Fonseca, J.P. et al. (2012). In vivo polycomb kinetics and mitotic chromatin binding distinguish stem cells from differentiated cells. Genes and Dev. 26: 857-871.

7. Coghill, J.M. et al. (2010). Separation of graft-versus-host dis-ease from graft-versus-leukemia responses by targeting CC-chemokine receptor 7 on donor T cells. Blood 115:4914-4922.

8. Rajan, S. et al. (2010). In vitro processing and secretion of mu-tant insulin proteins that cause permanent neonatal diabetes.


CELL SIGNALING & PROTEIN BIOLOGY Reporter Molecules

We offer a variety of tools for various reporter molecules:

Reporter Assays Reporter Stable Cell Lines Recombinant Fluorescent Proteins

Antibodies to Reporter Molecules Reporter Viral Vectors

Reporter Assays, Cell Lines and Reagents

GFP ELISA Kit

Recent Product Citations 1. Pfeiffer, B. et al. (2012). Using translational enhancers to in-

crease transgene expression in Drosophila. PNAS 109:6626-6631.

2. Wamboldt, Y. et al. (2009). Participation of leaky ribosome scan-ning in protein dual targeting by alternative translation initiation in higher plants. Plant Cell 21:157-167.


GFP ELISA Kit Colorimetric 96 Assays AKR-121

When direct quantitation of GFP fluorescence levels is desired, our GFP Quantitation Kit provides a su-perior method over time-consuming flow cytometry and semi-quantitative imaging techniques. This kit measures fluorescence levels directly in a plate-based fluorometer.

123

ß-Galactosidase Staining Kit

LacZ is a commonly used reporter gene in transfec-tion experiments because its gene product, ß-galactosidase, is extremely stable and resistant to proteolytic degradation, making it easy to assay. Our ß-Galactosidase Staining Kit provides an effi-cient, easy-to-use method to determine the transfec-tion efficiency of the LacZ gene.


ß-Galactosidase Staining Kit 75 Assays AKR-100

Cell Line Catalog Number

293/CFP AKR-270

293/GFP AKR-200

293/Luc AKR-230

Reporter Stable Cell Lines

Recent Product Citation Black, S.A. et al. (2008). TGFß1 stimulates connective tissue growth factor (CCN2/CTGF) expression in human gingival fibro-blasts through a RhoA-independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGFß1-induced CCN2/CTGF expression. J. Biol. Chem. 283:10835-10847.


Reporter Molecules CELL SIGNALING & PROTEIN BIOLOGY


A549/GFP AKR-209

BT-549/GFP AKR-255

ES-2/GFP AKR-206

HeLa/GFP AKR-213

MCF-7/GFP AKR-211

MCF-7/Luc AKR-234

MDA-MB-231/GFP AKR-201

MDA-MB-231/GFP-RFP AKR-221

MDA-MB-231/Luc AKR-231

293/YFP AKR-280

293T/GFP-Puro AKR-202



NIH3T3/GFP AKR-214

OVCA429/GFP AKR-212

OVCAR-5/RFP AKR-254

SKOV-3/GFP-Luc AKR-225

SKOV-3/Luc AKR-232

SKOV-3/RFP AKR-253

T47D/GFP AKR-208


MDA-MB-436/RFP AKR-252

MDA-MB-231/RFP AKR-251

RFP ELISA Kit

Recent Product Citations 1. Zhang, K. et al. (2014). Block-cell-

printing for live single-cell printing. PNAS 111:2948-2953. (AKR-201, AKR-211, AKR-252)

2. Zhang, W. et al. (2012). Microfluidics separation reveals the stem-cell-like deformability of tumor-initiating cells. PNAS 109:18707-18712. (AKR-211, AKR-252)

3. Weerakkody, D. et al. (2013). Family of pH (low) insertion peptides for tumor targeting. PNAS 110:5834-5839. (AKR-213)

Each cell line expresses one or more reporter molecules.


RFP ELISA Kit Colorimetric 96 Assays AKR-122

Sensitive: Detection limit of 150 pg/mL

Easy Quantification: Measure RFP levels in a standard 96-well microplate reader

Our RFP ELISA Kit provides a convenient, sensitive alternative to imaging systems and time-consuming FACS quantitation. The assay quantifies a wide vari-ety of red fluorescent protein variants including DsRed, TagRFP, TurboRFP, tdTomato, mCherry, mKate, mRuby, mBanana, mOrange, mPlum, and mStrawberry.

X-Gal Staining of Infected HUVEC Cells and Chick CAM Tissue. Left: HUVEC cells were infected with purified Ad-ß-Gal at 50 MOI (multiplicity of infection). X-gal staining was performed after 48 hour infection period. Right: Purified Ad-ß-Gal was injected intravenously into a 10-day old chick embryo. After three days, X-gal staining was performed on the chick chorioallanoic membrane tissue.

CELL SIGNALING & PROTEIN BIOLOGY Reporter Molecules


Monoclonal Antibodies to Reporter Molecules

Antibodies are provided at a concentration of 1 mg/mL. GFP antibody also recognizes EGFP, YFP, EYFP and CFP. RFP antibody recognizes Tag-RFP, Turbo-RFP, DeRed, mCherry and mOrange. Antibodies are suitable for Western blot, Immunostaining, ELISA and Dot blot.

Size Catalog Number

Mouse Anti-GFP Monoclonal Antibody (clone GF28R) 100 µg AKR-020

Mouse Anti-RFP Monoclonal Antibody (clone RF5R) 100 µg AKR-021

Product Name

Reporter Viral Vectors

Premade viruses and viral constructs with reporter molecules make ideal controls for viral gene delivery experi-ments.

Recombinant Fluorescent Proteins


Recombinant EGFP 100 µg STA-201

5 x 100 µg STA-201-5

100 µg STA-202

5 x 100 µg STA-202-5 Recombinant RFP

Recombinant EGFP and RFP are provided at a con-centration of 1 mg/mL and includes a 6xHis-tag at the C-terminus.

Recent Product Citation Sokolova, E. et al. (2013). Enhanced transcription rates in mem-brane-free protocells formed by coacervation of cell lysate. PNAS 110:11692-11697. (STA-201)

Recent Product Citation Maamary, J. et al. (2012). Attenuated influenza virus constructs with enhanced hemagglutinin protein expression. J. Virol. 86:5782-5790. (AKR-021)

Reporter AAV Reporter Adenoviruses

Reporter Lentiviruses

Reporter Retroviral Plasmids


AAV1-GFP Control Virus AAV-301

AAV2-Cre Control Virus AAV-310

AAV2-Luc Control Virus AAV-320






GFP Lentivirus Control LTV-300

RFP Lentivirus Control LTV-301


-Galactosidase ADV-002

Firefly Luciferase ADV-008

GFP ADV-004


GFP RTV-002

GFP RTV-051

Vector Backbone

pBABE

pMCs

GFP pMX RTV-050

GFP pMYs RTV-052

GFP-Puro pMX RTV-053

Epitope Tags/Antibodies CELL SIGNALING & PROTEIN BIOLOGY


Size Catalog Number

Mouse Anti-FLAG Tag Monoclonal Antibody (clone FG4R) 100 µg AKR-004

Mouse Anti-GST Tag Monoclonal Antibody (clone GST.B6) 100 µg AKR-005

Product Name

Mouse Anti-HA Tag Monoclonal Antibody (clone HA.C5) 100 µg AKR-006

Mouse Anti-His Tag Monoclonal Antibody (clone HIS.H8) 100 µg AKR-003

Mouse Anti-Myc Tag Monoclonal Antibody (clone Myc.A7) 100 µg AKR-007

Mouse Anti-V5 Tag Monoclonal Antibody (clone E10) 100 µg AKR-008

Mouse Anti-GAPDH Monoclonal Antibody 100 µg AKR-001

Mouse Anti-ß-Actin Monoclonal Antibody 100 µg AKR-002

Mouse Anti-ß-Tubulln Monoclonal Antibody 100 µg AKR-009

Monoclonal Antibodies to Epitope Tags

Antibodies are provided at a concentration of 1 mg/mL. GAPDH, ß-Actin and ß-Tubulin are also available as loading controls. All are suitable for Western blot, Immunostaining, ELISA, Immunoprecipitation, and Dot blot.

His-Tag Protein ELISA Kit

Our His-Tag Protein ELISA Kit allows you to detect and quantify His-tagged protein samples simply and reliably by comparing unknown samples with a recombinant standard. The kit is suitable for use with cell lysates and tissue homogenates.

Sensitive: Detect as little as 1 ng/mL protein or 50 pM of 6xHis-tag residues

Versatile: Use with proteins containing His-tag at either N- or C-terminus

Rapid Antibody Purification Kit


Rapid Antibody Purification Kit 10 Preps AKR-160

Species mg of IgG per Prep

Bovine 15-20

Goat 6-12

Human 20-30

Mouse 6-12

Rabbit 15-20

Our Rapid Antibody Purification Kit is designed for fast, single-step purification of high-quality IgG from ascites, serum, tissue culture media or hybridoma supernatants. IgG-containing samples are incubated with immobilized Protein A in the presence of a bind-ing buffer. Non-IgG components are washed and IgG is subsequently eluted. The supplied Protein A col-umn is suitable for 10 purification preps. The capac-ity per prep depends on the species of IgG; exam-ples are listed in the column at right. Capacity per Prep for the Rapid Antibody Purification Kit.

Recent Product Citation Dong, Y. et al. (2013). HMGB1 protein does not mediate the in-flammatory response in spontaneous spinal cord regeneration: A hint for CNS regeneration. J. Biol. Chem. 288:18204-18218.


His-Tag Protein ELISA Kit Colorimetric 96 Assays AKR-130

PhosphoBLOCKER™ Reagent

Dry Milk

PhosphoBLOCKER™ Western Blot Blocking Reagent

Superior Blocking with PhosphoBLOCKER™ Reagent. A549 cell lysate was blocked with dry milk or PhosphoBLOCKER before detection with anti-Phospho-p38 antibody.


1 L AKR-103

4 L AKR-104 PhosphoBLOCKER™ Western Blot Blocking Reagent


Phospho Antibody Stripping Solution 10 mL AKR-102

Phospho Antibody Stripping Solution

Western blot blockers such as dry milk or serum are sufficient to block unreactive sites on the membrane. However, they are not designed to preserve phos-phoprotein antigens during blotting.

High Sensitivity: Enhances low level phospho-protein signal without increasing background

Easy-to-use: Premixed dry blend

This solution removes anti-phosphoantibodies selec-tively from blots without significantly affecting the immobilized proteins, allowing re-probing of the blot with the same or a different antibody. Stripping of antibodies is done at room temperature, so no heat-ing of blots is required.

Multiple Blotting and Stripping of 4G10 Phosphotyrosine Antibody.

Recent Product Citations 1. Marques, J. et al. (2013). CRMP2 tethers kainate receptor activ-

ity to cytoskeleton dynamics during neuronal maturation. J. Neu-rosci. 33:18298-18310.

2. Ferreira, E. et al. (2013). Inflammatory cytokines induce a unique mineralizing phenotype in mesenchymal stem cells de-rived from human bone marrow. J. Biol. Chem. 288:29550-29561.

3. Rosich, L. et al. (2012). Counteracting autophagy overcomes resistance to everolimus in mantle cell lymphoma. Clin. Cancer Res. 18:5278-5289.

4. Martinez, P. et al. (2012). 53BP1 deficiency combined with te-lomere dysfunction activates ATR-dependent DNA damage response. J. Cell Biol. 197:283-300.

CELL SIGNALING & PROTEIN BIOLOGY Protein Phosphorylation



2 preps AKR-105

5 preps AKR-106 Phosphoprotein Purification Kit

Phosphoprotein Purification Kit

Enrichment of p-ERK. HeLa cell lysate was incubated with the Phos-phoprotein Enrichment Matrix from the Phos-phoprotein Purification Kit.

Our Phosphoprotein Purification Kit allows you to enrich your phosphoprotein samples quickly and easily. Phosphorylated proteins are affinity purified from lysates with a single-step purification / enrich-ment matrix. The entire procedure takes about 4 hours. Each prep can process 2.5 mg of total lysate protein, or approximately one confluent 10 cm dish.

Our Bacterial Protein Extraction Reagents contain a gentle, non-ionic detergent formulation that quickly extracts functional recombinant proteins from E. coli without mechanical disruption. The reagents are compatible with 6xHis and GST affinity purification systems and are suitable for small-scale or large-scale protein extraction.


Protein Isolation CELL SIGNALING & PROTEIN BIOLOGY

Rapid GST Inclusion Body Solubilization and Renaturation Kit


Rapid GST Inclusion Body Solubilization and Renaturation Kit 1 Kit AKR-110

The Rapid GST Inclusion Body Solubilization and Re-naturation Kit is designed to retrieve expressed GST fusion proteins in soluble form after lysis and extrac-tion. Each kit contains sufficient reagents to solubilize and renature up to 5-10 liters of bacterial culture.

Fast Results: No lengthy dialysis or dilution steps Easy-to-Use: No pH variation or redox pair Optimized: Designed specifically to solubilize and

renature GST inclusion bodies

Solubilization and Renaturation of GST-RTK Fusion Protein. Lane 1: MW STD; Lane 2: Whole E.Coli lysate; Lane 3, 7, 11: No detergent; Lane 4, 8, 12: 32-fold dilution; Lane 5, 9, 13: 8-fold dilution; Lane 6, 10, 14: 2-fold dilution.

1 2 3 4 5 6 7 8 9 10 11 12 13 14

Pre Beads Post Beads GS Beads

Recent Product Citations 1. Keller, D. et al. (2014). Mechanisms of HsSAS-6 assembly pro-

moting centriole formation in human cells. J. Cell Biol. 204:697-712.

2. Lalani, S. et al. (2013). MCTP2 is a dosage-sensitive gene re-quired for cardiac outflow tract development. Hum. Mol. Genet. 10.1093/hmg/ddt283.

Nuclear/Cytosolic Fractionation Kit


20 preps AKR-171

100 preps AKR-172 Nuclear/Cytosolic Fractionation Kit

The Nuclear/Cytosolic Fractionation Kit provides a simple and fast tool to isolate nuclear extract from the cytoplasmic fraction of mammalian cells. The opti-mized protocol provides high protein recovery and low cross-contamination in less than 2 hours. Ex-tracted fractions are functional and suitable for down-stream assays such as DNA footprinting, pre-mRNA splicing, gel shift assays, reporter assays, enzyme activity assays, and Western blot.

Size Catalog Number

5X Bacterial Protein Extraction Reagent (Phosphate) 50 mL AKR-181

5X Bacterial Protein Extraction Reagent (Tris) 50 mL AKR-180

Product Name

Bacterial Protein Extraction Reagents

HEK293 Cell Fractionation. Whole cell (W), cytosolic (C), and nuclear (N) frac-tions were im-munoblotted with Anti-Tubulin (cytosol specific) or Anti-Lamin A/C (nuclear specific).

CELL SIGNALING & PROTEIN BIOLOGY Protein Quantitation


High Sensitivity Protein Quantitation Assay Kit

Our High Sensitivity Protein Quantitation Assay Kit provides a robust method to quantify protein concen-trations in the µg/mL range. The fluorometric format creates a significant sensitivity advantage over stan-dard colorimetric methods such as Bradford or BCA assays.

0

50

100

150

200

250

0 100 200 300 400 500

BSA Standard (ug/mL)

RF

U

0

5

10

15

20

0 10 20 30 40

BSA Standard (ug/mL)

RF

U

Highly Sensitive: Quantify protein concentrations as low as 5 µg/mL

Fast: Obtain results in 5 to 15 minutes Easy-to-use: Read on a 96-well fluorescence

plate reader

Standard Curve Generated with the High Sensitivity Protein Quantitation Assay Kit.


High Sensitivity Protein Quantitation Assay Kit 100 Assays AKR-185

RIPA Buffer

RIPA buffer is a popular reagent for lysis of both adherent and suspension cells in culture, as well as making tissue homogenates. RIPA buffer extracts cytoplasmic, membrane, and nuclear proteins for a variety of down-stream protein assays and immunoassays. Our RIPA Buffer is provided as a 5X concentrate and is available with or without a Protease Inhibitor Cocktail.


5X RIPA Buffer 20 mL AKR-191

5X RIPA Buffer, with Protease Inhibitor Cocktail 20 mL AKR-190

METABOLISM RESEARCH Lipoprotein Metabolism


Lipoproteins are important assemblies of proteins and lipids that have implications in car-diovascular and related diseases. Our kits and reagents can help streamline the study of various members of lipoprotein metabolism pathways:

Lipoprotein Metabolism Research

Cholesterol / Lipoproteins Apolipoproteins Oxidized LDL Lipoprotein Receptors Cholesteryl Ester Pathway

Lipoprotein Lipase Triglycerides Free Fatty Acids Phospholipids Serum Proteins

Total Cholesterol Assay Kits



Fluorometric 192 Assays STA-390 Total Cholesterol Assay Kit

0

500

1000

1500

2000

2500

3000

3500

4000

4500

5000

1 to 10 1 to 100 1 to 1000 1 to 10000

RFUs

Serum and Plasma Dilutions

Plasma

Serum

Cholesterol Values from Normal Human Serum and Plasma Samples. Relative Fluorescence Unit (RFU) values are then com-pared against a Cholesterol Standard Curve (not shown).

Sensitive: Detect as little as 100 nM Fast: Simple 30 minute protocol Easy-to-use: Detect in a 96-well plate reader;

colorimetric and fluorometric formats available

Assay Principle for the Total Cholesterol Assay Kit (Fluorometric).

Cholesterol exists within lipoproteins in two forms: a free alcohol and a fatty cholesteryl ester, which is the predominant form of cholesterol transport and stor-age. Our Total Cholesterol Assay Kits provide a simple plate-based format that measures the amount of cho-lesterol in serum, plasma, cell lysates or tissue ho-mogenates. In the presence of cholesterol esterase, the assay will measure total cholesterol in both forms. In the absence of the esterase, the assay will meas-ure only free cholesterol. Quantitation is performed in a 96-well plate reader with your choice of colorimetric or fluorescence-based detection.

Recent Product Citations 1. Marino, A. et al. (2014). ITCH deficiency protects from diet-

induced obesity. Diabetes 63:550-561. (STA-384) 2. Mathews, E. et al. (2014). Mutation of 3-hydroxy-3-

methylglutaryl CoA synthase I reveals requirements for isopre-noid and cholesterol synthesis in oligodendrocyte migration arrest, axon wrapping, and myelin gene expression. J. Neurosci. 34:3402-3412. (STA-384)

Lipoprotein Metabolism METABOLISM RESEARCH


HDL and LDL/VLDL Cholesterol Assay Kit


HDL and LDL/VLDL Cholesterol Assay Kit Fluorometric 192 Assays STA-391

Our HDL and LDL/VLDL Cholesterol Assay Kit is similar in principle to our Total Cholesterol Assay Kit, but allows you to quantify HDL and LDL/VLDL sepa-rately in serum samples. After separating samples into HDL and LDL/VLDL fractions, the fluorometric assay is run according to the Assay Principle for the Total Cholesterol Assay Kit (previous page). In the presence of cholesterol esterase, the assay will measure total cholesterol in both free cholesterol and cholesteryl ester forms. In the absence of the es-terase, the assay will measure only free cholesterol. Quantitation of cholesteryl ester alone may be calcu-lated by subtracting free cholesterol from total choles-terol levels. Quantitation of Total Cholesterol, LDL/VLDL, and HDL.

7037

5011

1769

0

2000

4000

6000

8000

1

RFU

Total Cholesterol (HDL + LDL/VLDL)

LDL/VLDL

HDL

HDL-Cholesterol Assay Kit

Human Lipoproteins


Human High Density Lipoprotein (HDL) 100 µg STA-243

Human High Density Lipoprotein-2 (HDL-2) 100 µg STA-244

Human High Density Lipoprotein-3 (HDL-3) 100 µg STA-245

Human Low Density Lipoprotein (LDL) 100 µg STA-241

Human Low Density Lipoprotein (LDL), Copper (Cu++) Oxidized 100 µg STA-214

Human Low Density Lipoprotein (LDL), Malondialdehyde Modified 100 µg STA-212

Human Low Density Lipoprotein (LDL), Nitrated 100 µg STA-213

Human Very Low Density Lipoprotein (VLDL) 100 µg STA-242


HDL-Cholesterol Assay Kit Fluorometric 96 Assays STA-394

Sensitive: Detect as little as 1 µM Fast: Simple 45 minute assay protocol Easy-to-use: Detect in a 96-well plate fluores-

cence-based plate reader

Our HDL-Cholesterol Assay Kit provides a simple plate-based format similar to our Total Cholesterol Assays (previous page). This kit measures the amount of cholesterol in the HDL fraction isolated from serum, plasma, cell lysates or tissue homoge-nates.

Our human lipoproteins are isolated from human plasma following ultracentrifugation.




Human ApoAI ELISA Kit Colorimetric 96 Assays STA-362

Human ApoAII ELISA Kit Colorimetric 96 Assays STA-363

Human ApoCI ELISA Kit Colorimetric 96 Assays STA-364

Human ApoCII ELISA Kit Colorimetric 96 Assays STA-365

Human ApoCIII ELISA Kit Colorimetric 96 Assays STA-366

Human ApoE ELISA Kit Colorimetric 96 Assays STA-367

Human ApoB ELISA Kit Colorimetric 96 Assays STA-368

Human Apo(a) ELISA Kit Colorimetric 96 Assays STA-359

Human Apolipoprotein ELISA Kits

Apolipoproteins comprise the protein component of lipoprotein assemblies. Apolipoproteins fall into two classes: Non-exchangeable Apo B associates with LDL Exchangeable Apo A, C and E associate with

HDL Our Human Apo ELISA Kits provide a convenient and sensitive method for quantifying specific apolipopro-teins in serum, plasma, or other biological fluids.

Human ApoB ELISA Standard Curve. Detection Limits of Cell Biolabs’ Human Apo ELISA Kits.

Apo AI

Apo AII

Apo B

Apo CI

Apo CII

Apo CIII

Apo E

50 pg/mL

0.3 ng/mL

1 ng/mL

200 pg/mL

1 ng/mL

50 pg/mL

200 pg/mL

Apo (a)

1 ng/mL

Human Apolipoproteins


Human Albumin, Malondialdehyde Modified 100 µg STA-210

Human Apolipoprotein AI 100 µg STA-232

Human Apolipoprotein AII 100 µg STA-233

Human Apolipoprotein B-100 100 µg STA-234

Human Apolipoprotein B-100, Malondialdehyde Modified 100 µg STA-211

Human Apolipoprotein CI 100 µg STA-235

Human Apolipoprotein CIII 100 µg STA-237

Following ultracentrifugation, lipoproteins are isolated from human plasma. Water-soluble apolipoproteins are then purified from delipidated lipoprotein.



Human ApoAI and ApoB Duplex ELISA Kit


Human ApoAI and ApoB Duplex ELISA Kit Colorimetric 96 Assays STA-361

Efficient: Quantify two apolipoproteins from the same sample in just a few hours

Sensitive: Detect as little as 0.1 ng/mL of ApoAI and 1 ng/mL of ApoB from serum or plasma

Quantitative: Compare results to known ApoAI and ApoB standards

Human ApoAI and ApoB Duplex ELISA Kit Assay Principle.

As the primary protein components of HDL and LDL respectively, ApoAI and ApoB are arguably the most significant apolipoproteins in lipid metabolism re-search. Our Human ApoAI and ApoB Duplex ELISA Kit pro-vides a convenient tool to quantify both proteins in a single serum or plasma sample. Unlike other multi-plex assays, our ApoAI and ApoB Duplex ELISA does not require a luminometer for detection. Simply quantify both proteins using a standard colorimetric ELISA plate reader. The ELISA plate is coated with Anti-ApoAI and Anti-ApoB antibodies, which respectively capture HDL and LDL from the sample. Biotinylated Anti-ApoAI and Anti-ApoB detection antibodies are added, followed by enzyme conjugates. Alkaline phosphatase sub-strate is added allowing quantitation of ApoAI. After washing, HRP is added to allow quantiation of ApoB.


Sheep Anti-Human Apolipoprotein (a) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-131

Goat Anti-Human Apolipoprotein AI Polyclonal Antibody Immunoblot/ELISA 100 µg STA-132

Rabbit Anti-Human Apolipoprotein AII Polyclonal Antibody Immunoblot/ELISA 100 µg STA-133

Goat Anti-Human Apolipoprotein B-100/48 Polyclonal Antibody Immunoblot/ELISA 100 µg STA-134

Rabbit Anti-Human Apolipoprotein CI Polyclonal Antibody Immunoblot/ELISA 100 µg STA-135

Rabbit Anti-Human Apolipoprotein CII Polyclonal Antibody Immunoblot/ELISA 100 µg STA-136

Rabbit Anti-Human Apolipoprotein CIII Polyclonal Antibody Immunoblot/ELISA 100 µg STA-137

Goat Anti-Human Apolipoprotein E Polyclonal Antibody Immunoblot/ELISA 100 µg STA-138

Antibodies to Apolipoproteins

Antibodies are affinity purified.



OxiSelect™ Human Oxidized LDL ELISA Kits

LDL contains a hydrophobic core of various lipids surrounded by one molecule of Apolipoprotein B-100 (ApoB-100), which promotes solubility of the LDL in blood. LDL, often described as “bad” cholesterol, is even more dangerous when it becomes oxidized. Oxi-dized LDL (OxLDL) is more reactive with surrounding tissues and can collect within the inner lining of arter-ies. Our OxiSelect™ Human Oxidized LDL ELISA Kits are designed for the detection and quantitation of modi-fied LDL in human plasma or serum. Kits are avail-able to detect MDA-LDL, CML-LDL, or HNE-LDL in either the protein or lipid component of LDL. Our OxPL-LDL kit specifically detects oxidation in the phospholipid component of LDL.


OxiSelect™ Human Oxidized LDL ELISA Kit (CML-LDL Quantitation) Colorimetric 96 Assays STA-388

OxiSelect™ Human Oxidized LDL ELISA Kit (HNE-LDL Quantitation) Colorimetric 96 Assays STA-389

OxiSelect™ Human Oxidized LDL ELISA Kit (MDA-LDL Quantitation) Colorimetric 96 Assays STA-369

OxiSelect™ Human Oxidized LDL ELISA Kit (OxPL-LDL Quantitation) Colorimetric 96 Assays STA-358

Sensitive: Detect as little as 50 ng/mL of MDA-LDL, 150 ng/mL of CML-LDL, 150 ng/mL of HNE-LDL, or 100 ng/mL of OxPL-LDL

Quantitative: Compare unknown samples with provided copper oxidized LDL standard

Quantitation of MDA-LDL in Serum and Plasma Samples. Se-rum and plasma samples were treated with LDL Precipitation Solu-tion. Precipitated LDL pellets were resuspended in 1.6 mL of PBS before further dilution 1:160 in Assay Diluent according to the As-say Protocol. OxiSelect™ Human Oxidized LDL ELISA Assay Principle.

MDA is the most commonly found damage marker in oxidized LDL, but it can degrade in frozen samples after 1-2 months. CML and HNE, while less commonly found in OxLDL, may be more reliably detectable in samples that have been frozen for several months.



Human LDL Receptor ELISA Kit

Human LOX-1 ELISA Kit


Human LDLR ELISA Kit Colorimetric 96 Assays STA-386


Human LOX-1 ELISA Kit Colorimetric 96 Assays STA-387

Recent Product Citation Wu, J. et al. (2013). Clinical nephrology—IgA nephropathy, lupus nephritis, vasculitis. Nephrol. Dial. Transplant. 28:i175-i184.

Cholesterol can be toxic when accumulated in excess in cell membranes. The low-density lipoprotein recep-tor (LDLR) is the primary means of removing choles-terol from the circulation. LDLR is a transmembrane protein that transports cholesterol-carrying lipoprotein particles (primarily LDL) into cells. Receptor-ligand complexes enter the cell by endocytosis; bound lipo-proteins are subsequently released in the low-pH set-ting of the endosome, while the receptors return to the cell surface. Our Human LDLR ELISA Kit provides a simple, con-venient method for the detection and quantitation of LDL receptor in a variety of human sample types.

Sensitive: Detect as little as 50 pg/mL of human LDLR

Versatile: Assay is compatible with plasma, serum, cell lysates and tissue homogenates

Quantitative: Compare results to a known human LDLR standard

Standard Curve Generated with the Human LDLR ELISA Kit.

Standard Curve Generated with the Human LOX-1 ELISA Kit.

Monocytes and macrophages can form atheroscle-rotic lesions when they take in oxidized LDL (OxLDL). Uptake is done via the lectin-like oxidized LDL recep-tor-1 (LOX-1), which is expressed in vascular endo-thelium as well as in vascular smooth muscle cells, differentiated macrophages and platelets. LOX-1 can be cleaved and released as a soluble form (sLOX-1), which can serve as a prognostic biomarker in serum for early acute coronary syndromes, stroke and coro-nary heart disease. Our Human LOX-1 ELISA Kit provides a simple, con-venient method for the detection and quantitation of LOX-1 receptor in a variety of sample types.

Sensitive: Detect as little as 40 pg/mL of human LOX-1

Versatile: Assay is compatible with plasma, serum, cell lysates, tissue homogenates, or cell culture supernatants

Quantitative: Compare results to a known human LOX-1 standard

CETP Promotes Bidirectional Transfer of Cholesteryl Esters (CE) and Triglycerides (TG) Between Lipoproteins.



Human PCSK9 ELISA Kit


Human PCSK9 ELISA Kit Colorimetric 96 Assays STA-385

Standard Curve Generated with the Human PCSK9 ELISA Kit.

Proprotein convertase subtilisin kexin 9 (PCSK9) is a member of the proteinase K subfamiliy of subtilisin-related serine endoproteases. PCKS9 mediates LDL receptor (LDLR) degradation by binding to the EGF domain of the LDLR. This binding prevents LDLR from being sorted to the endosomes for recycling back to the cell surface. Instead, the PCSK9/LDLR complex is distributed to the lysosomes for degrada-tion. Our Human PCKS9 ELISA Kit provides a simple, convenient method for the detection and quantitation of PCSK9 in a variety of human sample types.

Sensitive: Detect as little as 150 pg/mL of human PCSK9

Versatile: Assay is compatible with plasma, serum, and cell and tissue lysates

Quantitative: Compare results to a known human PCSK9 standard

Human Cholesteryl Ester Transfer Protein (CETP) ELISA Kit


Human Cholesteryl Ester Transfer Protein (CETP) ELISA Kit Colorimetric 96 Assays STA-614

Cholesterol exists within lipoproteins in two forms: a free alcohol and a fatty cholesteryl ester. The choles-teryl ester is the predominant form of cholesterol transport and storage. Cholesteryl ester transfer protein (CETP) promotes the transfer of both cholesteryl esters and triglyc-erides between various types of lipoprotein particles: HDL, LDL, VLDL, and chylomicrons. Our Human CETP ELISA Kit provides a simple, con-venient method for the detection and quantitation in a variety of human sample types.

Sensitive: Detect as little as 60 ng/mL of human CETP

Versatile: Assay is compatible with plasma, serum, and other biological fluids

Quantitative: Compare results to a known human CETP standard



Lecithin Cholesterol Acyltransferase (LCAT) Activity Assay Kit

Lecithin Cholesterol Acyltransferase (LCAT) ELISA Kit


Lecithin Cholesterol Acyltransferase (LCAT) Activity Assay Kit Fluorometric 100 Assays STA-615


Lecithin Cholesterol Acyltransferase (LCAT) ELISA Kit Colorimetric 96 Assays STA-616

Lecithin cholesterol acyltransferase (LCAT) is an en-zyme that is associated with lipoproteins and plays a key role in promoting the transfer of excess cell-associated cholesterol from peripheral tissues to the liver to be excreted. LCAT catalyzes the transfer of an sn-2 acyl group from phosphatidylcholine to cholesterol, forming a cholesteryl ester. LCAT is bound to various lipopro-teins in the blood, including HDL and LDL. Our LCAT Activity Assay Kit provides a simple, con-venient method for measuring the phospholipase ac-tivity of LCAT in a variety of sample types including plasma, serum, cell lysates and tissue homogenates. Quantitation of LCAT activity is performed in a 96-well fluorescence-based plate reader. This assay may also be used to quantify other cal-cium independent phospholipase activities such as lipoprotein phospholipase A2 (LP-PLA2).

Assay Principle for the LCAT Activity Assay Kit. The close proximity of fluorescence labels on a dual-labeled fluorogenic probe keeps the fluorescence quenched. Upon cleavage of the probe by LCAT, fluorescence of the monomers can be measured at an excitation of 342 nm and emission of 400 nm.

Standard Curve Generated with the Lecithin Cholesterol Acyl-transferase (LCAT) ELISA Kit.

Lecithin cholesterol acyltransferase (LCAT) is an en-zyme that is associated with lipoproteins and plays a key role in promoting the transfer of excess cell-associated cholesterol from peripheral tissues to the liver to be excreted. LCAT catalyzes the transfer of an sn-2 acyl group from phosphatidylcholine to cholesterol, forming a cholesteryl ester. LCAT is bound to various lipopro-teins in the blood, including HDL and LDL. Our LCAT ELISA Assay Kit provides a simple, con-venient method for quantifying LCAT levels in a vari-ety of sample types including plasma, serum, and cell and tissue lysates. Quantitation of LCAT activity is performed in a standard 96-well plate reader.

Sensitive: Detect as little as 30 ng/mL of LCAT Versatile: Suitable for human, mouse, rat or rabbit

samples



Lipoprotein Lipase (LPL) Activity Assay Kit

Our Lipoprotein Lipase (LPL) Activity Assay Kit pro-vides a simple, convenient method to measure LPL activity in a variety of sample types. This kit uses a fluorogenic triglyceride analog as a lipase substrate. The quenched substrate is cleaved at the sn-1 posi-tion by LPL producing a fluorescent product that can be detected in a 96-well fluorescence plate reader. This assay will also measure the activity of endothe-lial and hepatic lipases. It cannot distinguish between these lipases and LPL.

Lipoprotein Lipase (LPL) ELISA Kit

Recent Product Citations 1. Dib, L. et al. (2014). LXRalpha fuels fatty acid-stimulated oxygen

consumption in white adipocytes. J. Lipid Res. 55:247-257. 2. Navab, M. et al. (2013). Transgenic 6F tomatoes act on the

small intestine to prevent systemic inflammation and dyslipide-mia caused by Western diet and intestinally derived lysophos-phatidic acid. J. Lipid Res. 54:3403-3418.


Lipoprotein Lipase (LPL) Activity Assay Kit Fluorometric 100 Assays STA-610


Lipoprotein Lipase (LPL) ELISA Kit Colorimetric 96 Assays STA-611

Assay Principle for the LPL Activity Assay Kit. The fluorogenic substrate is initially quenched and non-fluorescent. Upon cleavage of the probe by incubation with lipoprotein lipase (LPL), fluores-cence can be measured at an excitation of 342 nm and emission of 400 nm.

Lipoprotein lipase (LPL) is the key plasma lipase re-sponsible for the hydrolysis of the triglyceride core found in very low density lipoprotein (VLDL) particles formed in the liver. A mutation in the gene coding for LPL can lead to deficiencies in the enzyme, resulting in a diminished ability to breakdown fatty acids. Such LPL deficiency is known as chylomicronemia or Type I hyperlipoproteinemia. Our Lipoprotein Lipase (LPL) ELISA Kit provides a simple, convenient method to measure LPL levels in plasma, serum or other biological fluids from a variety of species (see below). LPL amounts are quantified against the provided LPL Standard in a colorimetric 96-well microplate reader.

Standard Curve Generated with the Lipoprotein Lipase (LPL) ELISA Kit.

Sensitive: Detect as little as 20 ng/mL of LPL Versatile: Suitable for human, rat, bovine, guinea

pig, or chicken samples (but not mouse)

Lipoprotein lipase (LPL) is the key plasma lipase responsible for the hydrolysis of the triglyceride core found in very low density lipoprotein (VLDL) particles formed in the liver.



Serum Triglyceride Quantitation Kits

Triglycerides serve as an energy source and play a key role in lipid metabolism. Lipases secreted into the intestines hydrolyze the triglyceride ester bond, pro-ducing glycerol and free fatty acids. Hepatic lipases also break down triglycerides in the liver to assemble very low density lipoprotein (VLDL) particles. Our Serum Triglyceride Quantitation Kits use a cou-pled enzymatic reaction system to measure triglyc-eride concentrations. First, a lipase hydrolyzes the ester bond, yielding free glycerol. The glycerol is then phosphorylated and oxidized, producing hydrogen peroxide, which reacts with the probe provided with each kit. Kits are available with either colorimetric or fluorescence-based detection, both of which are per-formed in a 96-well microtiter plate.

Standard Curve Generated with the Serum Triglyceride Quantitation Kit (Colorimetric).



Fluorometric 100 Assays STA-397 Serum Triglyceride Quantification Kit

Sensitive: Detect as little as 10 µM (1 mg/dL) with the colorimetric format and 2 µM (0.2 mg/dL) with the fluorometric format

Versatile: Suitable for serum, plasma, and cell and tissue lysates

Free Fatty Acid (FFA) Assay Kits

Free fatty acids (FFA) are released upon hydrolysis of triglycerides. FFAs then bind plasma albumin for circulation in the body, serving as a readily absorbed energy source for muscle, brain, and other organ tis-sues. Our Free Fatty Acid Assay Kits use a coupled enzy-matic reaction system to measure free fatty acid con-centrations in serum or plasma. Acyl CoA Synthetase catalyzes FFA acylation of CoA. The Acyl-CoA is then oxidized by Acyl CoA Oxidase, producing hydro-gen peroxide, which reacts with the kit’s probe. Kits are available with either colorimetric or fluorescence-based detection, both of which are performed in a 96-well microtiter plate.



Fluorometric 100 Assays STA-619 Free Fatty Acid Assay Kit

Standard Curve Generated with the Free Fatty Acid Assay Kit (Fluorometric).

Want to measure free glycerol content? See our Free Glycerol Assay Kits on page 146.

Recent Product Citations 1. Chellan, B. et al. (2014). IL-22 is induced by S100/calgranulin

and impairs cholesterol efflux in macrophages by downregulat-ing ABCG1. J. Lipid Res. 55:443-454. (STA-396)

2. Marino, A. et al. (2014). ITCH deficiency protects from diet-induced obesity. Diabetes 63:550-561. (STA-396)



Acetylcholine Assay Kits

Phosphatidylcholine Assay Kit

Sphingomyelin Assay Kit



Fluorometric 100 Assays STA-602 Acetylcholine Assay Kit

Our Acetylcholine Assay Kits provide a simple, convenient method to quantify acetylcholine in plasma, serum, cell suspensions, or tissue homogenates. Kits are available with either colorimetric or fluorometric detection, both of which are performed in a 96-well microplate reader.


Phosphatidylcholine Assay Kit Fluorometric 96 Assays STA-600

Phosphatidylcholine is the foremost phospholipid in eukaryotic cell membranes and comprises about 70% of the total phospholipids in plasma lipoproteins. Our Phosphatidylcholine Assay Kit is a simple fluoro-metric assay that measures phosphatidylcholine in plasma, serum, cell suspensions or tissue homoge-nates. Phospholipase D enzyme hydrolyzes phos-phatidylcholine into phosphatidic acid and choline. The choline is then oxidized by choline oxidase to produce hydrogen peroxide, which is detected by a fluorogenic probe in the presence of horseradish per-oxidase (HRP).


Sphingomyelin Assay Kit Fluorometric 96 Assays STA-601

0

500

1000

1500

2000

2500

3000

3500

0 5 10 15 20 25

RFU

s

Sphingomyelin (mg/dL)

Our Sphingomyelin Assay Kit is a simple fluorometric assay that measures sphingomyelin levels in plasma, serum, cell suspensions or tissue homogenates. Sphingomyelinase hydrolyzes sphingomyelin into ceramide and phosphocholine, which in turn is bro-ken down into choline. Choline is enzymatically oxi-dized to produce hydrogen peroxide, which is de-tected with a fluorogenic probe in the presence of horseradish peroxidase (HRP).

Standard Curve Generated with the Sphingomyelin Assay Kit.

Recent Product Citation Winklger, E.A. et al. (2014). Blood-spinal cord barrier disruption contributes to early motor-neuron degeneration in ALS-model mice. PNAS 111:E1035-E1042.



Human C-Reactive Protein ELISA Kit

Human Plasminogen ELISA Kit


Human C-Reactive Protein (CRP) ELISA Kit Colorimetric 96 Assays STA-392


Human Plasminogen ELISA Kit Colorimetric 96 Assays STA-393

Standard Curve Generated with the Human Plasminogen ELISA Kit.

Standard Curve Generated with the Human C-Reactive Protein (CRP) ELISA Kit.

C-Reactive Protein (CRP) is a serum protein that binds with high affinity to phosphocholine residues as well as other autologous and extrinsic ligands. CRP is a well-established marker of inflammation and tissue damage, and it has been associated with cardiovas-cular disease, atherosclerosis, and other diseases. Our Human C-Reactive Protein (CRP) ELISA Kit pro-vides a simple, convenient method to measure CRP levels in human plasma, serum, or other biological fluids. CRP amounts are quantified against the pro-vided CRP Standard in a colorimetric 96-well mi-croplate reader.

Sensitive: Detect as little as 1 ng/mL of CRP Versatile: Suitable for plasma, serum, or other

biological fluids

Plasminogen is a plasma glycoprotein that plays a role in macrophage recruitment, arterial stenosis, atherosclerosis, aneurysm formation, wound healing, and neovascularization. Plasminogen exists as an inactive proenzyme, but when converted to the active enzyme plasmin it serves to digest fibrin. This activa-tion is catalyzed by tissue-type plasminogen activator (tPA) or urokinase-type plasminogen activator (uPA). Our Human Plasminogen ELISA Kit provides a sim-ple, convenient method to measure plasminogen lev-els in human plasma, serum, or other biological flu-ids. Plasminogen amounts are quantified against the provided Plasminogen Standard in a colorimetric 96-well microplate reader.

Sensitive: Detect as little as 150 pg/mL of plas-minogen

Versatile: Suitable for plasma, serum, or other biological fluids




Human Albumin 100 µg STA-230

Human Albumin, Malondialdehyde Modified 100 µg STA-210

Human C-Reactive Protein 100 µg STA-240

Human Plasminogen 100 µg STA-239


Goat Anti-Human Albumin Polyclonal Antibody Immunoblot/ELISA 100 µg STA-130

Rabbit Anti-Human C-Reactive Protein Polyclonal Antibody Immunoblot/ELISA 100 µg STA-140

Goat Anti-Human Plasminogen Polyclonal Antibody Immunoblot/ELISA 100 µg STA-139

Human Serum Proteins

Antibodies to Human Serum Proteins

Antibodies are affinity purified.

Human Albumin ELISA Kit


Human Albumin ELISA Kit Colorimetric 96 Assays STA-383

Standard Curve Generated with the Human Albumin ELISA Kit.

Our human albumin and oxidized albumin were isolated and purified by HPLC. C-reactive protein was isolated from human pleural fluid. Plasminogen was isolated and purified from human plasma following an ultracentrifu-gation procedure.

Human serum albumin (HSA) is the most abundant protein in human plasma, constituting about half the protein in blood serum. It is typically found in concen-trations around 50 mg/mL. Produced in the liver in a preproalbumin state, albumin transports hormones, fatty acids, and other compounds through the circula-tion. It also maintains pH and osmotic pressure. Our Human Albumin ELISA Kit provides a simple, convenient method to measure HSA levels in human plasma, serum, urine, or other biological fluids. Hu-man albumin amounts are quantified against the pro-vided HSA Standard in a colorimetric 96-well mi-croplate reader.

Sensitive: Detect as little as 100 pg/mL of human serum albumin

Versatile: Suitable for plasma, serum, or other biological fluids



Lipid Extraction Kit, Chloroform Free

Lipid Quantification Kit


50 Preps STA-612 Lipid Extraction Kit (Chloroform Free)


Lipid Quantification Kit Colorimetric 100 Assays STA-613

Traditional methods of extracting lipids from tissues or cells, such as the well-published Folch method, have used chloroform as the extraction solvent. This method has a couple of disadvantages. First, the or-ganic phase ends up below the aqueous phase, cre-ating problematic removal through the upper phase that risks contamination. Second, chloroform has been classified in many places are a probable human carcinogen. Our Lipid Extraction Kit overcomes both disadvan-tages of the Folch method. The kit provides a chloro-form-free extraction method, and the use of proprie-tary organic solvents places the organic phase above the aqueous phase, allowing easy removal without disturbing the aqueous layer. Each kit provides suffi-cient reagents for 50 extractions from 100 µL sample sizes, but reagents may be scaled up for larger sam-ples.

Total Cholesterol Assay Performed on Extracted Lipids. Lipids extracted from fetal bovine serum (FBS) and HEK293 cells were prepared using the traditional Folch method (blue) and the Lipid Extraction Kit (red). Samples were tested for the presence of cho-lesterol in the Total Cholesterol Assay Kit (Cat. #STA-390).

Standard Curve Generated with the Total Lipid Quantification Kit.

Our Lipid Quantification Kit provides a convenient plate-based method to measure the total unsaturated lipid content found in various samples.* The assay uses a sulfo-phospho-vanillin method in which sam-ples are acidified and heated to solubilize and prime the lipids. The lipids then react with vanillin in acidic conditions to form a colorimetric product detectable in a standard 96-well microplate reader. This kit is compatible with plasma and serum sam-ples, or with crude or purified lipids extracted from cells. Each kit provides sufficient reagents for 100 assays including standards and unknown samples.

*Saturated lipids are not detected with this assay.


METABOLISM RESEARCH Renal Function Assays

Renal Function Assays

Our Renal Function Assays provide a simple, sensitive method to test for various markers related to kidney function:

Uric Acid / Uricase Urea

Creatinine Protein Carbamylation

Uric Acid/Uricase Assay Kit

Uric acid is the final oxidation end product of purine nucleotide metabolism. Uric acid is a potent antioxi-dant that is released during hypoxic conditions and is usually excreted in the urine via glomerular filtra-tion. In the urine, the enzyme uricase metabolizes uric acid to allantoin which is excreted from the body. Our Uric Acid / Uricase Assay Kit is a simple 96-well microplate-based assay for measuring concentra-tions of either uric acid or uricase in serum, plasma or urine samples. Detection is performed in a fluo-rescence-based plate reader.


Uric Acid/Uricase Assay Kit Fluorometric 400 Assays STA-375

Assay Principle for the Uric Acid / Uricase Assay Kit.


Urea Assay Kit Fluorometric 192 Assays STA-382

Sensitive: Detect as little as 0.5 µM of uric acid or 1 mU/mL of uricase

Quantitative: Kit includes both uric acid and uricase standards

Urea is the end product of protein nitrogen metabo-lism and is the primary vehicle for removing toxic ammonia from the body. Urea quantitation is one of the most widely applied tests for kidney function evaluation. Our Urea Assay Kit is a simple 96-well microplate-based assay for measuring urea concentrations in serum, plasma, lysates, or urine samples. Detection is performed in standard colorimetric plate reader.

Urea Assay Kit

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1 to 2 1 to 4 1 to 8

OD

63

0 n

m

Plasma and Serum Dilutions

Plasma

Serum

Human Plasma and Serum Samples Tested with the Urea Assay Kit.

Sensitive: Detect as little as 1 mg/dL of urea Quantitative: Measure unknown samples

against a known urea standard curve


Renal Function Assays METABOLISM RESEARCH

Urinary Creatinine Assay Kit


Urinary Creatinine Assay Kit Colorimetric 192 Assays STA-378

Creatinine is a metabolite formed from creatine and phosphocreatine (p-creatine). Subsequently creatinine enters the blood and is then excreted by the kidneys via glomerular filtration. Intra-individual variation of creatinine levels is <15% daily, making it a useful marker for normalizing levels of other mole-cules found in the urine. Our Urinary Creatinine Assay Kit is based on the Jaffe reaction between creatinine and alkaline picrate, which produces an orange-red color com-plex that can be easily read by a standard mi-croplate reader. A creatinine standard is provided to allow quantitative measurements of creatinine levels in urine samples. The assay is simple and takes less than one hour to perform.

Creatinine Levels in Urine Samples in the Presence and Absence of Glucose.

Protein Carbamylation ELISA Kit and Antibodies


OxiSelect™ Protein Carbamylation Sandwich ELISA Colorimetric 96 Assays STA-877

Goat Anti-Carbamyl-Lysine (CBL) Polyclonal Antibody Immunoblot/ELISA 50 µg STA-077

Rabbit Anti-Carbamyl-Lysine (CBL) Polyclonal Antibody Immunoblot/ELISA 50 µg STA-078

Carbamyl Lysine-BSA N/A 10 µg STA-379

Carbamylation is a post-translational modification which occurs throughout the lifespan of proteins in vivo. Carbamylation results from the binding of iso-cyanic acid, which spontaneously arises from high concentrations of urea, to lysine residues of proteins as carbamyl-lysine (CBL). Our Protein Carbamylation Sandwich ELISA Kit is a convenient microplate-based method for the evalua-tion of protein carbamylation in a variety of sample types. In addition, polyclonal antibodies are available for use in Western blot and ELISA applications.

Standard Curve Generated with the OxiSelect™ Protein Carbamylation Sandwich ELISA Kit.

Formation of Carbamyl-Lysine (CBL) During Carbamylation of Proteins.

METABOLISM RESEARCH Alcohol Assays


Alcohol Assay Kits

Our Alcohol Assay Kits provide a convenient plate-based method to measure primary alcohols, including ethanol, in plasma, serum or saliva samples.* The kit uses an enzymatic oxidation reaction that produces hydrogen peroxide, which reacts with the provided probe. Kits are available with either colorimetric or fluores-cence-based detection, both of which are performed in a 96-well microtiter plate. The colorimetric assay can detect alcohol levels as low as 30 µM, while the fluorometric assay detects as low as 15 µM.



Fluorometric 100 Assays STA-621 Alcohol Assay Kit

Free Glycerol Assay Kits



Fluorometric 100 Assays STA-399 Free Glycerol Assay Kit

Standard Curve Generated with the Free Glycerol Assay Kit.

Glycerol is the backbone of triglycerides. Lipases se-creted in the intestines hydrolyze the triglyceride ester bond, producing glycerol and free fatty acids. Our Free Glycerol Assay Kits use a coupled enzy-matic reaction system to measure free, endogenous glycerol concentrations. The glycerol is phosphory-lated and oxidized, producing hydrogen peroxide, which reacts with the probe provided with each kit. Kits are available with either colorimetric or fluores-cence-based detection, both of which are performed in a 96-well microtiter plate.

Standard Curve Generated with the Alcohol Assay Kit. *These assays are not suitable for urine samples.


PATHOGEN AND TOXIN ASSAYS Viral Core Antigens

Core antigens provide a convenient way to detect and quantify certain infectious viruses. We offer a variety of ELISA kits to measure the core antigen levels in plasma, serum, or purified virus preps. Note: Assays are for research use only, not for clinical diagnostic use.

Virus Core Antigen Detection

QuickTiter™ HIV-1 p24 ELISA Kit

A popular method of quantifying HIV-1 is the p24 ELISA. Our p24 ELISA kits provide a quick, convenient way to quantify the concentration of your lentivirus.


QuickTiter™ HIV Lentivirus Quantitation Kit (HIV-1 p24 ELISA) Colorimetric 96 Assays VPK-108-H

5 x 96 Assays VPK-108-H-5

QuickTiter™ MuLV Core Antigen ELISA Kit


QuickTiter™ MuLV Core Antigen ELISA Kit (MuLV p30) Colorimetric 96 Assays VPK-156

Sensitive: Detect as little as 300 pg/mL Fully quantitative: Recombinant core antigen in-

cluded as positive control

Murine leukemia virus (MuLV) is a retrovirus capable of causing cancer in mice and related vertebrates. Recently discovered in humans, xenotropic murine leukemia virus-related virus (XMRV) is closely related to MuLV; the p30 core antigens share 96% identity. Our QuickTiter™ MuLV Core Antigen ELISA Kit spe-cifically measures the level of the p30 core antigen in blood or other samples.

MuLV Core Antigen HIV-1 p24 Core Protein

Hepatitis B Core Antigen Hepatitis C Core Antigen

Standard Curve Generated with the QuickTiter HBV Core Antigen ELISA Kit.

0

0.5

1

1.5

2

2.5

3

0 5 10 15 20

MuL V p30 (ng /mL )

OD 450nm


QuickTiter™ HBV Core Antigen ELISA Kit

The QuickTiter™ HBV Core Antigen ELISA Kit spe-cifically quantifies the core protein of Hepatitis B vi-rus. A mouse monoclonal antibody is coated onto an 8 x 12 strip-well plate which allows the flexibility to save some of the wells for later use. The quantity of HBV is compared to the provided HBV Core Antigen Standard.

Sensitive: Detect as little as 1 ng/mL Fast: Results in about 5-6 hours Fully Quantitative: Recombinant core antigen

included as positive control


QuickTiter™ HBV Core Antigen ELISA Kit 96 Assays VPK-150

5 x 96 Assays VPK-150-5 Colorimetric

QuickTiter HBV Core Antigen ELISA Kit Standard Curve.

Viral Core Antigens PATHOGEN AND TOXIN ASSAYS

QuickTiter HCV Core Antigen ELISA Kit Standard Curve.


QuickTiter™ HCV Core Antigen ELISA Kit Colorimetric 96 Assays VPK-151

QuickTiter™ HCV Core Antigen ELISA Kit

Sensitive: Detect as little as 1 ng/mL Fast: Results in about 5-6 hours Fully Quantitative: Recombinant core antigen

included as positive control

The QuickTiter™ HBV Core Antigen ELISA Kit spe-cifically quantifies the core protein of Hepatitis B virus. A mouse monoclonal antibody is coated onto an 8 x 12 strip-well plate which allows the flexibility to save some of the wells for later use. The quantity of HBV is compared to the provided HBV Core Antigen Standard.

Recent Product Citation Chiu, C.Y. et al. (2014). Low-dose benzo(a)pyrene and its epox-ide metabolite inhibit myogenic differentiation in human skeletal muscle-derived progenitor cells. Toxicol. Sci. 10.1093/toxsci/kfu003. (STA-357)

Standard Curve Generated with the Aflatoxin Competitive ELISA Kit.


PATHOGEN AND TOXIN ASSAYS Toxin Assays

OxiSelect™ BPDE Adduct ELISA Kits

Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, ciga-rette smoke, and automotive exhaust. Benzo(a)pyrene, the first chemical carcinogen discovered, is converted to benzo(a)pyrene 7,8 diol-9,10 epoxide (BPDE) through a series of enzymatic reactions. BPDE can attack both proteins and DNA, forming adducts that can potentially result in tumor formation.


OxiSelect™ BPDE Protein Adduct ELISA Kit Colorimetric 96 Assays STA-301

OxiSelect™ BPDE DNA Adduct ELISA Kit Colorimetric 96 Assays STA-357

Standard Curve Generated Using the OxiSelect™ BPDE DNA Adduct ELISA Kit.

Our OxiSelect™ BPDE Adduct ELISA Kits provide a convenient method to measure BPDE adducts with either DNA or proteins from cells or tissues. Quantitation is performed by comparing unknown values against a standard curve generated with the standard provided in each kit.

Our BPDE DNA Adduct ELISA can detect adducts as low as 30 ng/mL.

Our BPDE Protein Adduct ELISA can detect adducts as low as 60 ng/mL.

Aflatoxin ELISA Kits

Aflatoxins are among the most potent genotoxic agents known. They can induce chromosomal ab-berations, micronuclei, sister chromatid exchange, unscheduled DNA synthesis, and chromosomal strand breaks. Aflatoxins can form adducts with both DNA and proteins. Our Aflatoxin Competitive ELISA Kits provide a con-venient method for detection of aflatoxin adducts. Our Aflatoxin Competitive ELISA Kit measures

the total of Aflatoxin B1 and Aflatoxin B2 adducts in protein samples.

Our Aflatoxin DNA Adduct Competitive ELISA Kit detects total Aflatoxin B1-DNA adducts, both ring-opened and ring-closed forms.


Aflatoxin DNA Adduct Competitive ELISA Kit Colorimetric 96 Assays AKR-351

Aflatoxin Competitive ELISA Kit Colorimetric 96 Assays AKR-350


Product Page 293 Cell Lines

AAV 47 Adenovirus 50 GFP Stable Expression 123 Lentivirus 59 Retrovirus 67

3-Nitrotyrosine Antibodies 85 ELISA Kit 85

4-HNE Antibodies 92 ELISA Kit 92

6-4PP Quantitation Kits 98 8-Iso-Prostaglandin F2

ELISA 92

8-Isoprostane ELISA Kit 92 8-Nitroguanine ELISA Kit 95 8-OHdG ELISA Kit 94 8-OHG ELISA Kit 95 A549/GFP Cell Line 123 AAV (Adeno-Assoc. Virus)

Cell Line 47 Expression Systems 41-45 Expression Vectors 46 Helper Free Systems 41-45 Packaging Systems 45 Premade Control Viruses 46 Purification Kits 47 Quantitation Kit 48 shRNA Expression 41-45 Titer Kit 48 Transduction Reagent 48

Acetylcholine Assays 140 Active Rac-GEF Assay 114 Adenovirus

Cell Line 50 Expression Systems 49, 81 microRNA Expression 81 Premade Recombinant 50-53 Purification Kits 54 Quantitation Kits 55 RCA Assay 56 shRNA Expression 49 Titer Kits 55 Transduction Reagent 56

Adhesion Assays 10-11 Adipogenesis Assay 27 Advanced Glycation End

Products Assay 88-89 Advanced Oxidation Protein

Products Assay 87 Aflatoxin Assays 150 AGE Assays 88-89 Albumin

Antibody 142 ELISA Kit 142 Protein 142

Product Page Apolipoproteins

Antibodies 133 ELISA Kits 132 Proteins 132

Arf1 Activation Assay 112-113

Arf6 Activation Assay 112-113

AUF1 Retroviral Vector 68 Autophagy Expression

Vectors 30

-Actin Antibody 125 -Galactosidase

Recombinant Adenovirus 50 Reporter Assays 123

ß-Tubulin Antibody 125 Bacterial Protein Extraction

Reagents 127

Biochips for Cell Adhesion 10 Blocking Reagent 126 BPDE

DNA Adduct ELISA 150 Protein Adduct ELISA 150

BrdU ELISA Kit 25 BT-549/GFP Cell Line 123 C3 Expression Vector 116 CA9 Recombinant

Adenovirus 50

c-Abl Retroviral Vector 67 cAMP ELISA Kits 119 Cancer Cell Assays

Angiogenesis 29 Anoikis 23 Cell Adhesion 10-11 Cell Invasion 18-19 Cell Migration 12-19 Cell Transformation 6-7 Colony Formation 6-9 Soft Agar 6-9 Tumor Cell Isolation Kit 9 Tumor Sensitivity 8

Carbamyl Lysine Antibodies 145 ELISA Kits 145

Carbonyl Assays 86 Carboxyethyl Lysine ELISA 88 Carboxymethyl Lysine

Assays 89

Catalase Activity Assays 106 Cdc42

Activation Assay 112-

Agarose Beads 115 Recombinant Adenovirus 51 Recombinant Protein 118 Retroviral Vector 69

Product Page Alcohol Assays 146 Aldehyde-Induced DNA

Damage Assays 99

Alkaline Phosphatase Assays 38 Angiogenesis

Recombinant Adenovirus 50 Tube Formation Assay 29

Anoikis Assays 23 Antibodies

Albumin 142 Apolipoproteins 133 Beta-Actin 125 Beta-Tubulin 125 Carboxymethyl Lysine (CML) 89 C-Reactive Protein 142 Flag Tag 125 Fluorescent Proteins 124 GAPDH 125 GFP 124 GST Tag 125 HA Tag 125 His Tag 125 HNE (4-Hydroxynonenal) 92 MDA (Malondialdehyde) 91 Methylglyoxal (MG) 89 Myc Tag 125 Nitrotyrosine 85 Plasminogen 142 RFP 124

Antibody Tools Blocking Reagent 126 Purification Kit 125 Western Stripping Solution 126

Antioxidant Assays Catalase Activity Assay 106 Cellular Antioxidant Asssay 109 Glutathione Assay 108 Glutathione Reductase Assay 108 HORAC Assay 110 ORAC Assay 110 Superoxide Dismutase

Activity Assay 107 Total Antioxidant Capacity

Assay 110 AOPP Assay 87 AP Sites Quantitation Kit 96 Apo(a) ELISA 132 ApoAI ELISA 132 ApoAI/ApoB Duplex ELISA 133 ApoAII ELISA 132 ApoB ELISA 132 ApoCI ELISA 132 ApoCII ELISA 132 ApoCIII ELISA 132 ApoE ELISA 132

PRODUCT INDEX


Product Page Cell Lines (cont’d)

MEF Feeder Cells 35 NIH3T3/GFP 123 OVCA429/GFP 123 OVCAR-5/RFP 123 Plat-A Retroviral Packaging

Cells 64

Plat-E Retroviral Packaging Cells 64

Plat-GP Retroviral Packaging Cells 64

SKOV-3/GFP-Luc 123 SKOV-3/Luc 123 SKOV-3/RFP 123 SNL Feeder Cells 35 T47D/GFP 123

Cell Migration Assays 12-19 Cell Proliferation Assays 24-25 Cell Transformation Assays 6-7 Cell Viability Assay 22 Cellular Antioxidant Assay 109 Cellular Senescence Assays 23 CETP ELISA Kit 136 cGMP ELISA Kits 119 Checkpoint Kinase Assays 121 Chemotaxis Assays 15, 19

Cholesterol Assays 130-131

Cholesteryl Ester Transfer Protein Assay

136

Clonogenic Tumor Cell Isolation Kit

9

CML (Carboxymethyl Lysine) Antibodies 89 Assays 89

CML-LDL ELISA Kit 93, 134

c-Myc Retroviral Vectors 69 Colony Formation Assays

Cell Transformation Assays 6-7 Hematopoietic Colony

Forming Cell Assay 36

Stem Cell Colony Assay 37 Tumor Cell Isolation Kit 9 Tumor Sensitivity Assay 8

Comet Assay Kits & Slides 97

Core Antigen Assays 148-

CPD Quantitation Kits 98 C-Reactive Protein

Antibody 142 ELISA 141 Protein 142

Cre Recombinant Adenovirus 50 Creatinine Assay 145 CSK Recombinant

Adenovirus 53

Product Page Cyclic AMP ELISA Kits 119 Cyclic GMP ELISA Kits 119 CytoSelect™ Cell-Based

Assays

Anoikis 23 Cell Adhesion 10-11 Cell Invasion 18-19 Cell Migration 12-19 Cell Transformation 6-7 Cell Viability 22 Chemotaxis 15, 19 Colony Formation 6-9 Cytotoxicity 22 Haptotaxis 16 Phagocytosis 28 Soft Agar 6-9 Transmigration 17 Tumor Sensitivity 8 Wound Healing 20

Cytoskeleton Regulation

Activation Assays 112-114

Adenoviruses 51 Expression Vectors 116 Retroviral Vectors 69

Cytotoxicity Assay 22 DCC Recombinant

Adenovirus 51

DNA Damage Assays 6-4PP Quantitation 98 8-Nitroguanine ELISA Kit 95 8-OHdG ELISA Kit 94 Aldehyde-Induced Damage 99 AP Sites Quantitation Kit 96 BPDE Adduct ELISA 99 Comet Assays 97 CPD Quantitation 98 Double-Strand Break Assay 96 UV Damage 98

DNA Methylation Assays 100 ECM (Extracellular Matrix)

Assays

Cell Adhesion Assays 10 Cell Invasion Assays 18-19 Tube Formation Assay 29

Endothelial Tube Assay 29 Epitope Tag Antibodies 125 ERK2

Recombinant Adenovirus 52 Retroviral Vector 68

ERK5 Recombinant Adenovirus

52

ES/EC Cells Alkaline Phosphatase Assays 38 Colony Formation Assays 37 Retroviral Expression

Systems 34

Product Page CEA Recombinant

Adenovirus 50

CEL ELISA Kit 88 Cell-Based Assays

Adhesion 10-11 Angiogenesis 29 Anoikis 23 Cell Contraction 29 Cell Viability 22 Chemotaxis 15, 19 Colony Formation 6-9 Cytotoxicity 22 Haptotaxis 16 Invasion 18-19 Migration 12-19 Phagocytosis 28 Proliferation 24-25 Senescence 23 Soft Agar 6-9 Transformation 6-7 Transmigration 17 Tumor Sensitivity 8 Wound Healing 20

Cell Cycle Adenoviruses 51 Anoikis Assay 23 Cell Viability Assay 22 Cytotoxicity Assay 22 Retroviral Vectors 67 Senescence Assays 23

Cell Fractionation Kits 127 Cell Invasion Assays 18-19 Cell Lines

293AAV 47 293AD 50 293LTV 59 293RTV 67 293/CFP 123 293/GFP 123 293/Luc 123 293/YFP 123 293T/GFP-Puro 123 A549/GFP 123 BT-549/GFP 123 ES-2/GFP 123 HeLa/GFP 123 JK1 Feeder Cells 35 MCF-7/GFP 123 MCF-7/Luc 123 MDA-MB-231/GFP 123 MDA-MB-231/GFP-RFP 123 MDA-MB-231/Luc 123 MDA-MB-231/RFP 123 MDA-MB-436/GFP 123 MDA-MB-436/RFP 123 MDA-MB-468/GFP 123

PRODUCT INDEX

153

Product Page ES-2/GFP Cell Line 123 Ethanol Assays 146 Exoenzyme C3 Expression

Vector 116 Extracellular Matrix Kits

Cell Adhesion Assays 10 Cell Invasion Assays 18-19 Tube Formation Assay 29

Feeder Cells 35 FFA Assay Kits 139 Firefly Luciferase Recom-

binant Adenovirus 50 Flag Tag Antibody 125 Free Fatty Acid Assays 139 Free Glycerol Assays 146 Fyn Recombinant Adenovirus 53 Gap Closure Migration

Assays 13 GAPDH Antibody 125 GEF (Guanine Exchange

Factors) Activation Assays 114 Agarose Beads 115

GFP Antibody 124 ELISA Kit 122 Lentiviral Vectors 59 Quantitation Kits 122 Recombinant Adenovirus 50 Recombinant Lentivirus 59 Recombinant Protein 124 Retroviral Vectors 67 Stable Cell Lines 123

GGA3 Agarose Beads 115 Global DNA Methylation

Assays 100 Glutathione Assay 108 Glutathione Reductase Assay 108 Glycerol Assays 146 Glycoaldehyde-BSA 88 GPCR Signaling Products 119 GST

Antibody 125 Inclusion Body Solubilization

and Renaturation Kit 127

GTPase Assay Kits 112-113

HA Tag Antibody 125 Haptotaxis Assays 16 HBV Core Antigen ELISA 149 HCV Core Antigen ELISA 149 HDL

Assay 131 Lipoprotein, Human 131

HEK 293 Cell Lines AAV 47 Adenovirus 50


Product Page HEK 293 Cell Lines (cont’d)

GFP Stable Expression 123 Lentivirus 59 Retrovirus 67

HeLa/GFP Cell Line 123 Hematopoietic Colony

Forming Cell Assay 36

Hepatitis B Core Antigen ELISA

149

Hepatitis C Core Antigen ELISA

149

HIF-1

Assays 26, 101

Recombinant Adenovirus 50 High Density Lipoprotein

Assay 131 Protein 131

His Tag Antibody 125 Protein ELISA 125

HIV-1 p24 ELISA Kits 60-61,

148 HNE

Antibodies 92 Assays 92

HNE-LDL Assay 93, 134

hnRNPA0 Retroviral Vector 68 HORAC Assay Kit 110 H-Ras

Activation Assay 112-113

Recombinant Protein 118 HuB Retroviral Vector 68 HuC Retroviral Vector 68 HuD Retroviral Vector 68 HuR Retroviral Vector 68 Hydrogen Peroxide Assays 104 Hydroxyl Radical Antioxi-

dant Capacity Assay 110

Hypoxia Assays 26,

IFN Recombinant Adenovirus 52 IB Recombinant Adenovirus 53 IKK Recombinant Adenovirus 53 IL-2 Recombinant Adenovirus 52 Immunoblot Blocking

Reagent 126

In Vitro Angiogenesis Assay 29 In Vitro Tumor Sensitivity

Assay 8

Inclusion Body Solubilization 127 Induced Pluripotent Stem

Cells

Lentiviral Vectors 33 Retroviral Packaging Cells 32 Retroviral Vectors 32-33

Product Page Invasion Assays 18-19 iPS Cell Reprogramming

Lentiviral Vectors 33 Retroviral Packaging Cells 32 Retroviral Vectors 32-33

JK1 Feeder Cells 35 JNK1


Klf4 Retroviral Vectors 69 KOSM Viral Vectors 33 K-Ras


Recombinant Protein 118 LC3 Expression Vectors 30 LCAT Assays 137 LDH Cytotoxicity Assay 22 LDL

Assays 131 Lipoprotein, Human 131 Oxidized 134

LDL Receptor Assay 135 Lecithin Cholesterol Acyl-

transferase Assays 137

Lentivirus Cell Line 59 Concentration & Purification

Kits 62

Control Plasmids 59 Expression Systems 57-58 Expression Vectors 59 Packaging Systems 58 Premade Control Viruses 59 Purification Kits 62 Quantitation Kits 60-61 shRNA Expression 57-58 Titer Kits 60-61 Transduction Kits 63

Leukocyte Assays Adhesion 11 Transmigration 17

Lin-28 Retroviral Vectors 69 Lipid Extraction Kit 143 Lipid Peroxidation Assays

8-Isoprostane ELISA 92 HNE Adduct ELISA 92 Malondialdehyde (MDA)

Assays 90-91

TBARS Assay 90 Lipid Quantification Assay 143 Lipoprotein Lipase Assays 138 Lipoproteins

Assays 130-133

Human 131-132

Oxidized 134

PRODUCT INDEX


Product Page ORAC Assay Kit 110 OVCA429/GFP Cell Line 123 OVCAR-5/RFP Cell Line 123 Oxidized LDL

Assay Kits 93, 134

Lipoprotein, Human 131 Oxidized Proteins 89 OxiSelect™ Oxidative

Stress / Damage Assays

Antioxidant Assays 106-110

DNA / RNA Damage Kits 94-100

Lipid Peroxidation Assays 90-93 Protein Oxidation Assays 85-89

ROS Assays 102-

OxLDL Assay 93,

OxPL Assay 134 Oxygen Radical Antioxidant

Capacity (ORAC) Assay 110

p24 ELISA Kits 60-61,

148 p38

Recombinant Adenovirus 52 Retroviral Vectors 68

p53 Recombinant Adenovirus 51 Retroviral Vectors 67

p68 RNA Helicase Adenovirus 51 PABP Retroviral Vector 68 PAK1

PBD Agarose Beads 115 Recombinant Adenovirus 51

PCNA ELISA Kit 25 PCSK9 ELISA Kit 136 Peroxide Detection Assays 104 Phagocytosis Assays 28 Phosphatidylcholine Assay 140 Phospho Antibody Stripping

Solution 126

PhosphoBLOCKER™ Western Blot Blocking Reagent

126

Phosphoproteins Antibody Stripping Solution 126 Blocking Reagent 126 Purification Kit 126

PI3K Retroviral Vector 68 PKC Recombinant

Adenovirus 53

Plasminogen Antibody 142 ELISA Kit 141 Protein, Human 142

Product Page LOX-1 Assay 135 LPL Assays 138 Luciferase

Recombinant Adenovirus 50 Reporter Cell Lines 123

Malondialdehyde Antibodies 91 Assays 90-91

MAP Kinase Signaling Recombinant Adenovirus 52 Retroviral Vectors 68

MAPKAPK2 Recombinant Adenovirus 52 Retroviral Vector 68

MCF-7/GFP Cell Line 123 MCF-7/Luc Cell Line 123 MDA (Malondialdehyde)

Antibodies 91 Assays 90-91

MDA-LDL Assay 93, 134

MDA-MB-231/GFP Cell Line 123 MDA-MB-231/Luc Cell Line 123 MDA-MB-231/RFP Cell Line 123 MDA-MB-436/GFP Cell Line 123 MDA-MB-436/RFP Cell Line 123 MDA-MB-468/GFP Cell Line 123 MEF Feeder Cells 35 MEK1


MEK5 Recombinant 52

MEKK1 Recombinant Adenovirus

52

MEKK3 Recombinant Adenovirus

52

Methylglyoxal (MG) Antibody 89 ELISA Kits 89

Microfluidic Biochips 10 microRNA Analysis

Adenoviral Expression System 81

Clone Collection 74-80 Control Vectors 80 Expression Vectors 74-80 Functional Reporter System 82 Precursor Clone Collection 74-80 Retroviral Expression Vector 81 Transduction Enhancer 82

Migration Assays 12-19 miRNA Analysis

Adenoviral Expression System 81

Clone Collection 74-80 Control Vectors 80

Product Page miRNA Analysis (cont’d)

Expression Vectors 74-80 Functional Reporter System 82 Precursor Clone Collection 74-80 Retroviral Expression Vector 81 Transduction Enhancer 82

MKK3 Recombinant Adenovirus 52 Retroviral Vector 68

MKK4 Recombinant Adenovirus

52

MKK6 Recombinant Adenovirus 52 Retroviral Vector 68

MKK7 Recombinant Adenovirus

52

MTT Cell Proliferation Assay 24 MuLV p30 Core Antigen

ELISA 148

Myc Tag Antibody 125 MyoD Recombinant

Adenovirus 51

Myogenin Recombinant Adenovirus

51

myr-Akt Retroviral Vectors 68 myr-Rac1

Recombinant Adenovirus 52 Retroviral Vectors 69

NANOG Retroviral Vectors 69 N-Carboxyethyl Lysine

ELISA 88

N-Carboxymethyl Lysine Antibody 89 Assay Kits 89

NFB Recombinant Adenoviruses

53

NIH3T3/GFP Cell Line 123 Nitrated LDL 131 Nitric Oxide Assays 105 Nitroguanine ELISA Kit 95 Nitrotyrosine

Antibodies 85 Assay Kits 85

NOD2 Recombinant Adenovirus

53

N-Ras


Recombinant Protein 118 Nuclear / Cytosolic Cell

Fractionation Kits 127

NY-ESO-1 Recombinant Adenovirus

50

Oct-3/4 Retroviral Vectors 69 OHdG ELISA Kit 94 OHG ELISA Kit 95

PRODUCT INDEX

Product Page Plat-A Retroviral Packaging

Cells 64 Plat-E Retroviral Packaging

Cells 64 Plat-GP Retroviral Packaging

Cells 64 Platinum Retroviral

Expression Expression Systems 65 Packaging Cell Lines 64

pMX Retroviral Vectors 66-69 PRAK


Proliferating Cell Nuclear Antigen Assay 25

Proliferation Assays 24-25 Protease Retroviral Vectors 69 Protein Extraction Reagents 127 Protein Oxidation Assays

Advanced Glycation End Products (AGE) 88-89

Advanced Oxidation Protein Products (AOPP) 87

BPDE Adduct 87 Carbonyl 86 CEL (Carboxyethyl Lysine) 88 CML (Carboxymethyl Lysine) 89 MG (Methylglyoxal) 89 Nitrotyrosine 85

Protein Phosphorylation Antibody Stripping Solution 126 Blocking Reagent 126 Purification Kit 126

Protein Quantitation Kit 128 Proteins

Albumin 142 Apolipoproteins 132 EGFP 124 GRP-PH Domain 119 Oxidized/Nitrated 131

Purification Kits AAV 47 Adenovirus 56 Antibodies 125 Lentivirus 62 Phosphoproteins 126 Retrovirus 70

QuickTiter™ Viral Titer & Quantitation Kits AAV 48 Adenovirus 55 HBV Core Antigen 149 HCV Core Antigen 149 HIV p24 148 Lentivirus, Recombinant 60-61

Product Page

Reporter Genes Lentiviral Vectors 59

Quantitation Assays 122-123

Recombinant Adenovirus 50 Recombinant Lentivirus 59 Retroviral Vectors 67 Stable Cell Lines 123

Retrovirus Concentration & Purification

Kits 70

Expression Systems 65 Expression Vectors 66 Gene-Specific Vectors 67-69 Packaging Cell Lines 64, 67 Purification Kits 70 Quantitation Kits 71 shRNA Expression 66 Transduction Kits 72

RFP Antibody 124 ELISA Kit 123 Recombinant Protein 124 Stable Cell Lines 123

RGS Recombinant Proteins 118 Rho


Agarose Beads 115 Recombinant Adenovirus 51 Recombinant Proteins 118 Retroviral Vector 69

RhoA Activation Assay 112-113

RhoB Activation Assay 112-113

RhoC Activation Assay 112-113

Rho Kinase Activity Assays 120 RIPA Buffer 128 RNA Damage ELISA Kit 95 RNAi Enhancer Reagent 82 ROCK Activity Assay Kits 120

ROS Assays 102-105

scAAV Control Vector 46 Expression Systems 42-45 Expression Vector 46

SCGE Assay Kits 97 SEAP Recombinant

Adenovirus 50

Senescence Assays 23 shAkt Recombinant

Adenovirus 53

Renal Function Assays 144-


Product Page

QuickTiter™ Viral Titer &

MuLV p30 148 Retrovirus, Recombinant 71

Rab Recombinant Proteins 118 Rac


Agarose Beads 115 GEF Assay 114 Recombinant Adenovirus 51 Recombinant Proteins 118 Retroviral Vectors 69

Radius™ Cell Migration Assays

13

Raf1 Recombinant Adenovirus 52 Retroviral Vectors 68

Ral


Agarose Beads 115 Recombinant Proteins 118

Ran


Agarose Beads 115 Recombinant Protein 118

Rap


Recombinant Proteins 118 RAPAd® Adenoviral

Expression Systems 49

Rapid GST Inclusion Body Solubilization and Renatu-ration Kit

127

Rapid RCA Assay 56 Ras Superfamily


Agarose Beads 115 Expression Vectors 116 Recombinant Adenovirus 51 Recombinant Proteins 118 Retroviral Vectors 69

RCA Assay Kit 56 Reactive Oxygen Species

(ROS) Assays 102-105

Recombinant Adenoviruses 50-53 Recombinant Proteins

Fluorescent Proteins 124 GRP-PH Domain 119 Small GTPase 118

Rel B Recombinant Adenovirus

53

PRODUCT INDEX


Product Page shRNA Expression

Adeno-Associated Virus 41-45 Adenovirus 49 Lentivirus 57-59 Retrovirus 66

Single Cell Gel Electro-phoresis Assays

97

SKOV-3/GFP-Luc Cell Line 123 SKOV-3/Luc Cell Line 123 SKOV-3/RFP Cell Line 123 Small GTPase

Activation Assays 112-

Active GEF Assays 114 Agarose Beads 115 Expression Vectors 116 Premade Adenoviruses 51 Retroviral Vectors 69

SNL Feeder Cells 35 SOD Activity Assay Kit 107 Soft Agar Colony Assay Kits

Cell Transformation Assays 6-7

Hematopoietic Colony 36

Stem Cell Colony Formation 37

Tumor Cell Isolation Kit 9 Tumor Sensitivity Assay 8

SOK Recombinant 52

Sox2 Retroviral Vector 69 Sphingomyelin Assay 140 Src Recombinant Adenovirus 53 Stat5 Retroviral Vectors 68 Stem Cell Research

Alkaline Phosphatase Detection Kits 38

Feeder Cells 35

Hematopoietic Colony 36

iPS Cell Reprogramming 32-33 PCR Primers 38

Retroviral Expression 34

Stem Cell Colony Assay 37 Total Protein: ES Cell Line D3 38 Total RNA: ES Cell Line D3 38

Superoxide Dismutase Assay 107 T47D/GFP Cell Line 123 TAC Assay 110

Tac-Rac1 Recombinant Adenovirus

52

TBARS Assay Kit 90 TIA1 Retroviral Vector 68 Tiam1 Assay Kit 114 TIAR Retroviral Vector 68

Product Page

Total Antioxidant Capacity 110

Total Cholesterol Assays 130 Total Protein: ES Cell Line D3 38 Total RNA: ES Cell Line D3 38 Transcription Regulation

Retroviral Vectors 68 Transformation Assays 6-7 Transmigration Assays 17 Triglyceride Assays 139 TTP Retroviral Vector 68 Tube Formation Assay 29 Tumor Antigen Adenoviruses 50 Tumor Cell Assays

Cell Adhesion 10-11 Cell Invasion 18-19 Cell Migration 12-19 Cell Transformation 6-7 Chemosensitivity 8 Soft Agar Colony Formation 6-9 Transmigration 17 Tumor Cell Isolation 9

Tyrosine Kinase Adenoviruses 53

uPA / uPAR Retroviral Vectors 69

Urea Assay 144 Uric Acid / Uricase Asay 144 UV DNA Damage Assays 98 V5 Tag Antibody 125 VEGF Recombinant

Adenovirus 50 Very Low Density Lipoprotein

Assay 131 Lipoprotein, Human 131

ViraBind™ Purification Kits AAV 47 Adenovirus 54 Lentivirus 62 Retrovirus 70

ViraDuctin™ Transduction Kits & Reagents AAV 48 Adenovirus 56 Lentivirus 63 Retrovirus 72

Viral Expression Systems AAV 41-45 Adenovirus 49, 81 Lentivirus 57-58 Retrovirus 65

Viral Packaging Cells AAV 47 Adenovirus 50 Lentivirus 59 Retrovirus 64, 67

Product Page Viral Titer Kits

AAV 48 Adenovirus 55 Lentivirus 60-61 Retrovirus 71

Viral Transduction Reagents AAV 48 Adenovirus 56 Lentivirus 63 Retrovirus 72

ViraSafe™ Lentivirus Expression Systems 57-58

Virus Core Antigen Assays HBVcAg 149 HCVcAg 149 HIV-1 p24 148 MuLV p30 148

Virus Purification Kits AAV 47 Adenovirus 54 Lentivirus 62 Retrovirus 70

Virus Quantitation Kits AAV 48 Adenovirus 55 Lentivirus 60-61 Retrovirus 71

VLDL Assay 131 Lipoprotein, Human 131

VSV-G Retroviral Vector 67 Western Blot Blocking

Reagent 126

Wound Healing Assay 20 WST-1 Cell Proliferation

Reagent 24

PRODUCT INDEX

Please locate the contact information below for the office in your country. If your country is not listed, please contact Cell Biolabs’ Corporate Office at [email protected].

ALBANIA MEDICAL BIOMICS LTD. Tel: 0030 6974711158 [email protected]

ALGERIA KARAWAN SCIENTIFIC Tel: +216 71 230 933 Fax: +216 71 230 787 [email protected]

ARGENTINA GENBIOTECH S.R.L. Tel: +54 011-45561600 [email protected]

AUSTRALIA JOMAR BIOSCIENCE PTY LTD Tel: +61 8 8431 2041 Fax: +61 8 8431 2049 [email protected]

AUSTRIA THP MEDICAL PRODUCTS VERTRIEBS GMBH Tel: +43 1 292 82 80 Fax: +43 1 292 82 80-88 [email protected]

BIOCAT GMBH Tel: +49 (0) 6221 7141516 Fax: +49 (0) 6221 7141529 [email protected]

BAHRAIN DPC LEBANON S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected]

BELGIUM BIO-CONNECT BV Tel: +31 (0) 26 326 4450 Fax: +31 (0) 26 326 4451 [email protected]

BRAZIL SELLEX (S.A.C.) Tel: (011) 5506-4646 Fax: (011) 5505-7433 [email protected]

BULGARIA RIDACOM LTD. Tel: 003592 955 99 98 [email protected]

CHILE FERMELO S.A. Tel: +56 2 247 2976 Fax: +56 2 247 2977 [email protected]

CHINA BEIJING SEAJET SCIENTIFIC CO., LTD. (BEIJING OFFICE) Tel: +86 10 8859-7838 Fax: +86 10 8859-5011 [email protected]

BEIJING SEAJET SCIENTIFIC CO., LTD. (SHANGHAI OFFICE) Tel: +86 021 63599871 Fax: +86 021 63599873 [email protected]

GENETIMES TECHNOLOGY, INC Tel: +86 21 3367-6611 Fax: +86 21 3367-6155 [email protected]

COLOMBIA SUMINISTROS CLINICOS ISLA LTDA Tel: 571 6250580 [email protected]

COSTA RICA BIOCIENTIFICA INTERNACIONAL S. DE R.L. Tel: 506 2272-3700 Fax: 506 2272-0460 [email protected] www.biocientifica.net

CROATIA INTEROS2000 LTD. Tel: +385 1 617 0059 Fax: +385 1 617 0086 [email protected]

CZECH REPUBLIC SCINTILA, S.R.O. Tel: +420 567 302 681 Fax: +420 567 330 277 [email protected] www.scintila.cz

THP MEDICAL PRODUCTS VERTRIEBS GMBH Tel: +43 1 292 82 80 Fax: +43 1 292 82 80-88 [email protected]

DENMARK BIO-MEDIATOR KY Tel: +358-9-852 4898 Fax: +358-9-852 4884 [email protected] www.bio.fi

EGYPT NEW TEST CO. (NTCO) FOR SCIENTIFIC SERVICES Tel: +20 3 544 4736 Fax: +20 3 359 6836 [email protected]

ESTONIA UPSTREAM OU Tel: +372 51 86 295 [email protected] www.upstream.ee

FINLAND BIO-MEDIATOR KY Tel: +358-9-852 4898 Fax: +358-9-852 4884 [email protected] www.bio.fi

FRANCE EUROMEDEX Tel: +33 3 88 18 07 27 Fax: +33 3 88 18 07 28 [email protected] www.euromedex.com

www.cellbiolabs.com [email protected] 157

CANADA CEDARLANE LABORATORIES Tel: 1 289 288-0001 Fax: 1 289 288-0020 [email protected]

CYPRUS MEDICAL BIOMICS LTD. Tel: 0030 6974711158 [email protected]

HONG KONG BEIJING SEAJET SCIENTIFIC CO., LTD. [email protected]

GENETIMES TECHNOLOGY INTERNATIONAL HOLDING LTD Tel: 852 2385 2818 Fax: 852 2385 1308 [email protected]

HUNGARY THP MEDICAL PRODUCTS VERTRIEBS GMBH Tel: +43 1 292 82 80 Fax: +43 1 292 82 80-88 [email protected]

GREECE MEDICAL BIOMICS LTD. Tel: 0030 6974711158 [email protected]

GERMANY BIOCAT GMBH Tel: +49 (0) 6221 7141516 Fax: +49 (0) 6221 7141529 [email protected] www.biocat.com

GHANA THRIVUS COMPANY LTD Tel: +233-54-349-5200 Fax: +233-27-219-0060 [email protected] www.thrivus.com

WORLDWIDE CONTACTS

WORLDWIDE CONTACTS


IVORY COAST THRIVUS COMPANY LTD Tel: +233-54-349-5200 Fax: +233-27-219-0060 [email protected] www.thrivus.com

JAPAN COSMO BIO CO., LTD. Tel: +81 3 5632 9610 Fax: +81 3 5632 9614 [email protected] www.cosmobio.co.jp

JORDAN GENETICS COMPANY “EL WERATHA” Tel: +962 6 5536402 Fax: +962 6 5536398 [email protected] www.genetics-jo.com

DPC LEBANON S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected]

KOREA CMI BIOTECH Tel: 82-2-444-7101 Fax: 82-2-444-7201 [email protected] www.cmibio.com KOMA BIOTECH INC. Tel: 82-2-2660-5660 Fax: 82-2-2660-5669 [email protected] www.komabiotech.co.kr

KUWAIT DPC LEBANON S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected]

LATVIA ADRONA SIA Tel: +371 67551894 Fax: +371 67551976 [email protected] www.adrona.lv

LITHUANIA UPSTREAM OU Tel: +372 51 86 295 [email protected] www.upstream.ee

UPSTREAM OU Tel: +372 51 86 295 [email protected] www.upstream.ee

LUXEMBOURG BIO-CONNECT BV Tel: +31 (0) 26 326 4450 Fax: +31 (0) 26 326 4451 [email protected]

MACEDONIA MEDICAL BIOMICS LTD. Tel: 0030 6974711158 [email protected]

MALAYSIA AXON SCIENTIFIC SDN. BHD. Tel: +603 89451482 Fax: +603 89419421 [email protected]

MALTA KARAWAN SCIENTIFIC Tel: +216 71 230 933 Fax: +216 71 230 787 [email protected]

MEXICO PROBIOTEK Tel: +81 8383 4892 Fax: +81 8383 8907 [email protected]

MOROCCO KARAWAN SCIENTIFIC Tel: +216 71 230 933 Fax: +216 71 230 787 [email protected]

NETHERLANDS BIO-CONNECT BV Tel: +31 (0) 26 326 4450 Fax: +31 (0) 26 326 4451 [email protected]

NICARAGUA BIOCIENTIFICA INTERNACIONAL S. DE R.L. Tel: 506 2272-3700 Fax: 506 2272-0460 [email protected] www.biocientifica.net

NIGERIA THRIVUS COMPANY LTD Tel: +233-54-349-5200 Fax: +233-27-219-0060 [email protected] www.thrivus.com

NORWAY BIO-MEDIATOR KY Tel: +358-9-852 4898 Fax: +358-9-852 4884 [email protected] www.bio.fi

OMAN BIO MEDICAL SCIENTIFIC SERVICES LLC Tel: +971 3 762 0880 Fax: +971 3 762 0882 [email protected] www.biomss.com


PAKISTAN FY SCIENTIFIC ASSOCIATES Tel: +92 21 34327418 Fax: +92 21 3453798 [email protected] www.fybreeds.com MADINA SCIENTIFIC STORES Tel: +92 42 7238770 Fax: +92 42 7314122 [email protected] www.madinasci.com

PANAMA BIOCIENTIFICA INTERNACIONAL S. DE R.L. Tel: 506 2272-3700 Fax: 506 2272-0460 [email protected] www.biocientifica.net

ITALY VALTER OCCHIENA S.R.L. Tel: +39 011 7716971 Fax: +39 011 7761800 [email protected] www.valterocchiena.com

LEBANON DPC LEBANON S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected]

NEW ZEALAND JOMAR BIOSCIENCE PTY LTD Tel: +61 8 8431 2041 Fax: +61 8 8431 2049 [email protected]

PERU BIOLAB ANDINA SAC Tel: +51-1-4491223 Fax: +51-1-4491223 [email protected] www.biolabandina.com

INDIA BIOGENUIX MEDSYSTEMS PVT. LTD. Tel: +91-11-4875 4875 Fax: +91-11-2561 2008 [email protected] www.biogenuix.com

INDONESIA CV. KRISTALINDO BIOLAB Tel: +62 31 5998626 Fax: +62 31 5998627 [email protected]

IRAQ DPC LEBANON S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected]

IRELAND CAMBRIDGE BIOSCIENCE LTD. Tel: +44 (0) 1223 316 855 Fax: +44 (0) 1954 781 323 [email protected] www.bioscience.co.uk

ISRAEL BIOCONSULT Tel: +972-2-5667043 Fax: +972-2-5662790 [email protected] www.bioconsult.co.il

WORLDWIDE CONTACTS

www.cellbiolabs.com [email protected] 159

SLOVAKIA SCINTILA, S.R.O. Tel: +420 567 302 681 Fax: +420 567 330 277 [email protected] www.scintila.cz

THP MEDICAL PRODUCTS VERTRIEBS GMBH Tel: +43 1 292 82 80 Fax: +43 1 292 82 80-88 [email protected]

SLOVENIA THP MEDICAL PRODUCTS VERTRIEBS GMBH Tel: +43 1 292 82 80 Fax: +43 1 292 82 80-88 [email protected] www.thp.at

SOUTH AFRICA BIOCOM BIOTECH Tel: +27 12 644 2795 Fax: +27 86 654 9048 [email protected] www.biocombiotech.co.za

SPAIN BIONOVA CIENTIFICA S.L. Tel: +34 91 551 54 03 Fax: +34 91 433 45 45 [email protected] www.bionova.es

SRI LANKA BIOGENUIX MEDSYSTEMS PVT. LTD. Tel: +91-11-4875 4875 Fax: +91-11-2561 2008 [email protected] www.biogenuix.com

SWEDEN BIO-MEDIATOR KY Tel: +358-9-852 4898 Fax: +358-9-852 4884 [email protected] www.bio.fi

SWITZERLAND LUBIOSCIENCE GMBH Tel: +41 41 417 02 80 Fax: +41 41 417 02 89 [email protected] www.lubio.ch

BIOCAT GMBH Tel: +49 (0) 6221 7141516 Fax: +49 (0) 6221 7141529 [email protected]

TAIWAN GENETEKS BIOSCIENCE, INC. Tel: 886-2-8993 6968 Fax: 886-2-8993 6967 [email protected] HOHO TECHNOLOGY CO., LTD. Tel: 886 02 2748 9697 Fax: 886 2 8192 4210 E-mail: [email protected] www.hohoteco.idv.tw

THAILAND ADVANCED MEDICAL SCIENCE CO., LTD. Tel: +66-2-196-2066 Fax: +66-2-196-2069 [email protected] www.amsthailand.com

TUNISIA KARAWAN SCIENTIFIC Tel: +216 71 230 933 Fax: +216 71 230 787 [email protected]

TURKEY MEDSANTEK LTD. CO. Tel: +90 212 635 85 46 Fax: +90 212 635 83 50 [email protected] www.medsantek.com.tr

UKRAINE ALMABION LTD. Tel: +7 (473) 273-48-28 [email protected] www.almabion.ru

UNITED ARAB

BIO MEDICAL SCIENTIFIC SERVICES LLC Tel: +971 3 762 0880 Fax: +971 3 762 0882 [email protected] www.biomss.com

UNITED KINGDOM CAMBRIDGE BIOSCIENCE LTD. Tel: +44 (0) 1223 316 855 Fax: +44 (0) 1954 781 323 [email protected] www.bioscience.co.uk

URUGUAY GENBIOTECH S.R.L. Tel: +54 011-45561600 [email protected]

VIET NAM TRUONG BIO, INC. Tel: 84 8 3755 3648 Fax: 84 8 3755 4403 [email protected] www.truongbio.com UNITED SCIENTIFIC CO., LTD. Tel: +84 8 22446300 Fax: +84 8 39106816 [email protected] www.unitedscientific.net

UNITED STATES CELL BIOLABS, INC. Tel: +1 858 271-6500 Fax: +1 858 271-6514 [email protected] www.cellbiolabs.com

YEMEN BIOTEC MEDICAL CORP Tel: 00967 1 297680 [email protected]

SINGAPORE PRECISION TECHNOLOGIES PTE LTD. Tel: +65 6273 4573 Fax: +65 6273 8898 [email protected] www.pretech.com.sg

POLAND STI Tel: +48 (61) 6417759 Fax: +48 (61) 6417758 [email protected] www.sti.biz.pl

PORTUGAL MEDITECNO, LDA. Tel: +351 214 581 723 Fax: +351 214 077 945 [email protected] www.meditecno.pt

QATAR DPC LEBANON S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected]

ROMANIA TAXON SOLUTIONS SRL Tel: +40 729 852168 Fax: +40 21 2103500 [email protected]

RUSSIA ALMABION LTD. Tel: +7 (473) 273-48-28 [email protected] www.almabion.ru

SAUDI ARABIA SALIMA TRADING CORP. Tel: +966 1 493 0759 Fax: +966 1 493 2819 [email protected]


SERBIA PROMEDIA DOO. Tel: +381 11 344 6208 Fax: +381 11 344 6209 [email protected] www.promedia.rs

SWISS CONCEPT LTD. Tel: +381 11 3610 945 Fax: +381 11 3610 948


catalogo cell biolabs

Documents

cellbased assays

kinase assays reporter

protein biology

stem cell research

technical support

pricing kit components

us tollfree fax

metabolism research