2010 cell biolabs catalog final

CREATING SOLUTIONS FOR LIFE SCIENCE RESEARCH

CELL-BASED ASSAYS MIGRATION INVASION TRANSFORMATION ADHESION ANGIOGENESIS PHAGOCYTOSIS

CELL SIGNALING SMALL GTPASE RHO KINASE

PHOSPHORYLATION GPCR SIGNALING

STEM CELL RESEARCH IPS CELL REPROGRAMMING FEEDER CELLS

COLONY FORMATION ASSAYS ALK PHOS ASSAYS

Cover Images: Neurite Formation of Differentiated N1E-115 Neuroblastoma Cells. Images courtesy of Dr. Richard L. Klemke, University of California, San Diego.

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

VIRAL EXPRESSION ADENOVIRUS ADENO-ASSOCIATED VIRUS

LENTIVIRUS RETROVIRUS

OXIDATIVE & CELLULAR STRESS LIPID PEROXIDATION PROTEIN OXIDATION

DNA DAMAGE ANTIOXIDANTS ROS

Cell-Based Assays

3 www.cellbiolabs.com [email protected]

Ordering Information 4

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Stem Cell Research 23

Viral Expression 31

Oxidative & Cellular Stress 71

Cell Signaling 85

Product Index

Worldwide Distributors

94

98

TABLE OF CONTENTS

microRNA Analysis 61

ORDERING INFORMATION & SUPPORT

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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 nearly 60 countries. If you don’t see your country listed, please contact our U.S. office.

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

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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 96 Wells CBA-130

5 x 96 Wells 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. Kang, M-I. et al. (2009). A selective small-molecule nuclear

factor-kB inhibitor from a high-throughput cell-based assay for "activator protein-1 hits". Mol. Cancer Ther. 8:571-581.

2. Li, H. et al. (2009). Lysophosphatidic acid stimulates cell migra-tion, invasion, and colony formation as well as tumorigenesis/metastasis of mouse ovarian cancer in immunocompetent mice. Mol. Cancer Ther. 8:1692-1701.

3. Takezawa, K. et al. (2009). Sorafenib inhibits non-small cell lung cancer cell growth by targeting B-RAF in KRAS wild-type cells and C-RAF in KRAS mutant cells. Cancer Res. 69:6515-6521.

4. Lee, K.B. et al. (2008). Low energy proton beam induces tumor cell apoptosis through reactive oxygen species and activation of caspases. Exp. and Mol. Medicine 40(1):118-129.

5. Shen, L. et al. (2008). E1A inhibits the proliferation of human cervical cancer cells (HeLa cells) by apoptosis induction through activation of HER-2/Neu/Caspase-3 pathway. Med. Oncol. 25:222-228.

6. Wei, Q. et al. (2008). Sulfiredoxin is an AP-1 target gene that is required for transformation and shows elevated expression in human skin malignancies. PNAS 105:19738-19743.

7. Gazin, C. et al. (2007). An elaborate pathway required for Ras-mediated epigenetic silencing. Nature 449(7165):1073-1077.


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.

Easy Fluorescence Detection with the CytoSelect™ Cell Trans-formation Assay. HeLa and NIH3T3 cells were seeded at various concentrations and cultured for 6 days. Transformed colonies were quantified according to the assay protocol.

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*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

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Recent Product Citations 1. Hirata, H. et al. (2010). Role of secreted Frizzled-related pro-

tein3 in human renal cell carcinoma. Cancer Res. 70:1896-1905. (CBA-135)

2. 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)

3. Ke, X-S. et al. (2008). Epithelial to mesenchymal transition of a primary prostate cell line with switches of cell adhesion mod-ules but without malignant transformation. PLoS One 3(10): E3368. (CBA-135)

www.cellbiolabs.com [email protected]


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

5 x 96 Wells CBA-150-5

CytoSelect™ 96-Well In Vitro Tumor Sensitivity Assay

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Inhibition of HeLa Cell Transformation by 5-Fluorouracil. HeLa cells were seeded at 5000 cells/well and cultured 7 days at various 5-FU concentrations. Cell transformation was determined accord-ing to the assay protocol. IC50 value of 5-Fluorouracil on HeLa cell anchorage-independent growth was determined to be ~ 1 μM.

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 col-ony 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



CytoSelect™ Clonogenic Tumor Cell Isolation Kit

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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™ 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 Wells CBA-070


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


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


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


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


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


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

The CytoSelect™ ECM Cell Adhesion Assays provide a quantitative method for evaluation of cell adhesion. The 48-well plate is precoated with your choice of substrate. Cells are seeded onto the substrate; adher-ent 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

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CELL-BASED ASSAYS Cell Adhesion

BSA

Vitronectin

Laminin

Fibronectin

Collagen IV

Collagen I

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.

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


CytoSelect™ Leukocyte Endothelium Adhesion Assays




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

CytoSelect™ Leukocyte-Endothelium Adhesion Assay

CytoSelect™ Leukocyte-Epithelium Adhesion Assay

CytoSelect™ Leukocyte-endothelium Adhesion Assay Principle.

Human Monocytic THP-1 Adhesion to HUVEC Monolayer Us-ing the CytoSelect™ Leukocyte-endothelium Adhesion Assay. HUVEC monolayer in a 48-well plate was treated with 1 µM PMA for 12 hrs. LeukoTracker™ labeled THP-1 cells were allowed to attach to HUVEC monolayer for 1 hr. Adherent cells were lysed and quantified as described in the assay protocol.

Leukocyte interactions with vascular endothelium consist of a cascade of processes including the firm adhesion of leukocytes to endothelial cell adhesion molecules. The CytoSelect™ Leukocyte Endothelium Adhesion As-says provide a robust system for the quantitative determination of interactions between leukocytes and endo-thelium. Adherent cells can be quantified on a fluorescence plate reader.

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Human Breast Cancer MDA-231 Adhesion to HUVEC Monolayer Using the CytoSelect™ Tumor-endothelium Adhesion Assay. Left: CytoTracker™-labeled MDA-231 cells were allowed to attach to HUVEC monolayer in a 48-well plate for one hour. Adherent cells were lysed and quantified as described in the assay protocol. Right: HUVEC monolayer in 48-well plate was treated with 50 ng/ml TNFα for 12 hours. CytoTracker™-labeled MDA-231 cells (50,000 cells/well) were allowed to attach to HUVEC monolayer for one hour. Adherent cells were lysed and quantified as described in the assay protocol.

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CELL-BASED ASSAYS Cell Adhesion

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Recent Product Citation Xue, J. et al. (2009). NFkB regulates thrombin-induced ICAM-1 gene expression in cooperation with NFAT by binding to the intronic NFkB site in the ICAM-1 gene. Physiol. Genomics 38:42-53. (CBA-210)


CELL-BASED ASSAYS Cell Migration / Invasion

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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 and invasion assays use precision-molded plastic inserts, with and with-out coatings, to enable the study of a wide variety of migratory and invasive cells. All kits include dyes for full cell quantitation; no manual cell counting is required. Cell migration may take various forms. Use the following table to help select the best as-says for you based on your research goal and cell type.

Cell Migration and Invasion Assay Selection Guide

Assay Definition Cell

Types Pore Size

Insert Coating

Assay Formats

Chemotaxis

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

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 Migration of cells through the

vascular endothelium toward a chemoattractant

Leukocytes 3 µm None 24-Well

Tumor Cells 8 µm None 24-Well

Wound Healing Migration of cells across a “wound” gap Any N/A None 24-Well

Invasion

Movement of cells through the 3D extracellular matrix into

neighboring tissues; includes ECM degradation and

proteolysis


Tumor Cells

ECM Matrix (top)

24-Well, 96-Well

Collagen I (top)

24-Well, 96-Well

Laminin I (top)

24-Well, 96-Well

8 µm


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CytoSelect™ Cell Migration Assays—Chemotaxis

Product Name Pore Size Detection Size Catalog Number

CytoSelect™ 24-Well Cell Migration Assay

3 µm Fluorometric 12 Wells CBA-103


8 µm Colorimetric 12 Wells CBA-100


12 µm Colorimetric 12 Wells CBA-107





CytoSelect™ 96-Well Cell Migration Assay

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.

Assay Principle for the CytoSelect™ Cell Migration Assay.

10% FBS 0% FBS


Recent Product Citations 1. Awasthi, N. et al. (2009). Endothelial monocyte activating poly-

peptide II interferes with VEGF-induced proangiogenic signaling. Laboratory Investigation 89(1):38-46. (CBA-100)

2. Tabata, C. et al. (2009). All-trans-retinoic acid inhibits tumor growth of malignant pleural mesothelioma in mice. Eur. Respir. J. 34:1159-1167. (CBA-100)

3. Igarashi, J. et al. (2009). Transforming growth factor-ß1 down-regulates caveolin-1 expression and enhances sphingosine 1-phosphate signaling in cultured vascular endothelial cells. Am. J. Physiol. Cell Physiol. 297:C1263-C1274. (CBA-100)

4. 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)

5. Shynlova, O. et al. (2008). Monocyte chemoattractant protein-1 (CCL-2) integrates mechanical and endocrine signals that medi-ate term and preterm labor. J. Immunol. 181:1470-1479. (CBA-102)

6. Chatterjee, S. et al. (2009). Site-specific carboxypeptidase B1 tyrosine nitration and pathophysiological implications following its physical association with nitric oxide synthase-3 in experimental sepsis. J. Immunol. 183:4055-4066. (CBA-104)

7. Caglayan, E. et al. (2008). Differential roles of cardiomyocyte and macrophage peroxisome proliferator-activated receptor gamma in cardiac fibrosis. Diabetes 57:2470-2479. (CBA-105)

8. Beattie, A.J. et al. (2008). Chemoattraction of progenitor cells by remodeling extracellular matrix scaffolds. Tissue Eng. Part A 10.1089/ten.tea.2008.0162. (CBA-106)



CytoSelect™ Cell Migration Assays—Haptotaxis


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

Fluorometric 12 Wells CBA-101-COL

Colorimetric 12 Wells CBA-100-FN

Fluorometric 12 Wells 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™ Cell Migration Assays (previous page) or the Leukocyte Transmigration Assay (next page).

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Recent Product Citation Kamiya, K. et al. (2007). Protein Kinase C delta activated adhesion 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



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CytoSelect™ Cell Migration Assays—Transmigration


CytoSelect™ Leukocyte Transmigration Assay Fluorometric 24 Wells CBA-212

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

Pore Size

3 µm

8 µm

Cancer cell transmigration, particularly extravasa-tion, is an important step in cancer metastasis. The CytoSelect™ Cell Transmigration Assays provide a robust system for the quantitation of transmigrations and interactions 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. Yang, H. and H.E. Grossniklaus (2010). Constitutive overexpres-

sion of pigment epithelium derived factor inhibition of ocular mela noma growth and metastasis. Invest. Ophthalmol. Vis. Sci. 51:28-34. (CBA-216)

3. Liu, K. et al (2008). Lentivirus mediated gene transfer of PEDF results in decreased uveal melanoma transendothelial migration. Invest. Opthalmol. Vis. Sci 48:5244. (CBA-216)

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Assay Principle for the CytoSelect™ Leukocyte Transmigration Assay.

The Leukocyte Adhesion and Transmigration Cascade.

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.

16


Recent Product Citations 1. Hirata, H. et al. (2010). Role of secreted Frizzled-related pro-

tein3 in human renal cell carcinoma. Cancer Res. 70:1896-1905. (CBA-110)

2. Zhang, Q. et al. (2009). Nuclear factor-kB mediated Transform-ing Growth Factor-ß-induced expression of vimentin is an inde-pendent predictor of biochemical recurrence after radial prostatectomy. Clin. Cancer Res. 15:3557-3567. (CBA-110)

3. Cabello, C. et al. (2009). The cinnamon-derived Michael accep-tor cinnamic aldehyde impairs melanoma cell proliferation, inva-siveness, and tumor growth. Free Rad. Biol. and Med. 46(2):220-231. (CBA-110)

4. Ji, H. et al (2007). LKB1 modulates lung cancer differentiation and metastasis. Nature 48:807-810. (CBA-110, CBA-111, CBA-112)

5. Eckstein, N. et al. (2009). Hyperactivation of the insulin-like growth factor receptor I signaling pathway is an essential event for cisplatin resistance of ovarian cancer cells. Cancer Res. 69:2996-3003. (CBA-112)

6. Lam, K.K.W. et al. (2009). Glycodelin-A as a modulator of tro-phoblast invasion. Hum. Reprod. 24:2093-2103. (CBA-112)

7. Neil, J.R. et al. (2008). Cox-2 inactivates Smad signaling and enhances EMT stimulated by TGFß through a PGE2-dependent mechanism. Carcinogenesis 29:2227-2235. (CBA-112)

8. Thal, D.R. et al. (2008). Expression of coronin-3 (coronin-1C) in diffuse gliomas is related to malignancy. J. Pathol. 214:415-424. (CBA-112)


CytoSelect™ Cell Invasion Assays, continued


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


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

Fluorometric 12 Wells CBA-111-COL

Colorimetric 12 Wells CBA-110-LN

Fluorometric 12 Wells CBA-111-LN

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

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

CytoSelect™ 96-Well Cell Invasion Assay, Laminin I Fluorometric 96 Wells 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.

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NIH3T3 HT-1080

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Recent Product Citations 1. Liu, S. et al. (2008). MMP-activated anthrax lethal toxin demonstrates high potency in targeting tumor vasculature. J. Biol. Chem. 283: 529

-540. (CBA-100-C) 2. 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) 3. Alfano, R.W. et al. (2009). Matrix metalloproteinase-activated anthrax lethal toxin inhibits endothelial invasion and neovasculature forma-

tion during in vitro morphogenesis. Mol. Cancer Res. 7:452-461. (CBA-106-C) 4. Eckstein, N. et al (2008). EGFR-pathway analysis identified amphiregulin as a key factor for cisplatin resistance of human breast cancer

cells. J. Biol. Chem. 283:739-750. (CBA-106-C)

Product Name Pore Size Detection Size Catalog Number

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

Fluorometric 2 x 12 Wells CBA-101-C

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

8 µm


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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

Wound healing assays are useful for studying tissue matrix remodeling, regulation of cytoskeletal struc-ture, and cell proliferation and migration rates of dif-ferent cells and culture conditions. Traditional wound healing assays are performed by making a scratch across a confluent cell monolayer to create an open gap, mimicking a “wound”. Such scratch assays, however, lack a consistently defined wound area and result in high inter-sample variation. Our CytoSelect™ 24-Well Wound Healing Assay provides a more consistent method to measure cell migration across a “wound field” gap in vitro. Pro-prietary treated inserts generate a consistently de-fined 0.9mm gap between the cells. Cells can then be treated and monitored for proliferation or migra-tion across the wound field by imaging samples at fixed time points or time-lapse microscopy.


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

5 x 24 Wells 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.

Assay Principle for the CytoSelect™ 24-well Wound Healing Assay.

0%

50%

100%

Percent Wound Closure


19

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 (Red Blood Cell) Colorimetric 96 Assays CBA-220

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

CELL-BASED ASSAYS Phagocytosis

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

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

Recent Product Citations 1. Dowling, D.J. et al. (2010). 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)

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


Assay Principle for the CytoSelect™ 96-Well Phagocytosis Assay (Red Blood Cell).

CELL-BASED ASSAYS Cell Viability / Death

20

CytoSelect™ Anoikis Assays


CytoSelect™ 24-Well Anoikis Assay Colorimetric / Fluorometric

24 Wells CBA-080

CytoSelect™ 96-Well Anoikis Assay Colorimetric / Fluorometric

96 Wells 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.

This assay allows you to quantify and monitor anoikis in cells using a Poly-HEMA precoated plate. Live cells can be viewed under a microscope and quanti-fied on a plate reader by MTT (colorimetric) or Cal-cein AM (fluorometric), both included with the kit. Dead cells are detected with 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

CytoSelect™ Cell Viability and Cytotoxicity Assay


CytoSelect™ Cell Viability and Cytotoxicity Assay Kit Colorimetric / Fluorometric

1 plate* CBA-240

Cell viability characteristics include cellular meta-bolic activity and cell membrane integrity. Our Cy-toSelect™ Cell Viability and Cytotoxicity Assay pro-vides both a colorimetric and fluorometric format for monitoring cell viability via metabolic activity. Live cells are detected with MTT (colorimetric detec-tion) 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 com-pounds or agents that affect cell viability. This kit is suitable for eukaryotic cells, not bacteria or yeast.

*Each kit provides sufficient reagent quantities to perform 24 or 96 assays in a 24-well or 96-well plate respectively.

Recent Product Citations 1. 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) 2. Liu, H. et al (2008). Cysteine-rich protein 61 and connective tissue growth factor induce de-adhesion and anoikis of retinal pericytes.

Endocrinology 149:1666-1677. (CBA-080)

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 live and dead cells by micros-copy, 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


Cellular Senescence Assays


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

96-Well Cellular Senescence Assay (SA β-gal Activity) Fluorometric 96 Wells CBA-231

Senescence Associated (SA) β-galactosidase is a common biochemical marker of cellular senescence. Cells expressing such markers have been identified in vivo in tissues. Our Cellular Senescence Assays provide an efficient method to detect senescent cells. SA ß-Gal catalyzes hydrolysis of X-gal, which produces a blue color in senescent cells. The results may be visualized using our SA β-gal Staining Kit. If higher throughput is desired, quantify senescent cells on a fluorometric plate reader using our 96-Well Cellu-lar Senescence Activity Assay. This kit uses a fluorometric substrate to detect SA ß-Gal activity.

Cell Contraction Assay (Collagen-Based)

CELL-BASED ASSAYS Senescence, Cell Contraction


Cell Contraction Assay Light Microscopy 24 Assays CBA-201

Assay Principle for the Collagen-Based Cell Contraction Assay.

0

1

2

3

4

5

6

0 50 100 150

Time (min)

Co

ntr

ac

tio

n (

mm

)

Control

BDM

Wound healing is comprised of epithelialization, connective tissue deposition, and contraction. The contraction process is believed to be mediated by specialized fibroblasts (myofibroblasts). 3D collagen gels have been widely used in fibroblast contraction studies.

Contraction Inhibition by BDM. 5 x 105 COS-7 cells in 0.5 mL collagen gel lattice were cultured for 2 days. Before initiation of contraction, cells were pretreated with 10mM BDM for 1 hour. The change in gel diameter (mm) was measured with a ruler at various times following release.

21 www.cellbiolabs.com [email protected]

Recent Product Citation 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. Endocrinol-ogy 151:12571268. (CBA-201)

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.

Endothelial Tube Formation (In Vitro Angiogenesis) Assay

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.

For angiogenesis to occur, endothelial cells must escape their stable location and break through the base-ment membrane. Cells migrate toward an angiogenic stimulus that may be released from nearby tumor cells. These cells proliferate to form new blood vessels.


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

CELL-BASED ASSAYS Angiogenesis, Autophagy

22

Recent Product Citations 1. Weskamp, G. et al. (2010). Pathological neovascularization is

reduced by inactivation of ADAM17 in endothelial cells but not in pericytes. Circ. Res. 10.1161/CIRCRESAHA.109.207415. (CBA-200)

2. Alfano, R.W. et al. (2009). Matrix metalloproteinase-activated anthrax lethal toxin inhibits endothelial invasion and neovascula-ture formation during in vitro morphogenesis. Mol. Cancer Res. 7:452-461. (CBA-200)

3. Nogueras, S. et al (2008). Coupling of endothelial injury and repair. An analysis using an in vivo experimental model. Am. J. Physiol. Heart Circ. Physiol. 294:H708-H713. (CBA-200)

4. Masamune, A. et al (2008). Hypoxia stimulates pancreatic stel-late cells to induce fibrosis and angiogenesis in pancreatic can-cer. Am. J. Physiol. Gastrointest. Liver Physiol. 295:G709-G717. (CBA-200)

GFP-LC3 Expression Vectors

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.


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

Our Endothelial Tube Formation Assay provides an easy, robust system to assess angiogenesis in vitro. The assay uses an ECM gel matrix de-rived from mouse sarcoma cells; this matrix very closely resembles an in vivo basement mem-brane environment.

pCMV-GFP-LC3 Vector Map.


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.

24

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 page 57.

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

Lentiviral Polycistronic Vector for iPS Cell Generation

STEM CELL RESEARCH

25


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

pLenti-p53 shRNA (Mouse) Lentiviral Vector 100 µL LTV-451

Our pLentG-KOSM Lentiviral Vector provides a con-venient 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 four genes are con-trolled by a single CMV promoter. The transcription factor ORF is followed by IRES-GFP as a reporter to verify viral transduction into your target cell. Efficien-cies of iPS generation are typically higher compared to transduction of four separate viruses each contain-ing a single gene. p53 shRNA has recently been shown to potentially increase efficiency of iPS production. Our p53 shRNA lentiviral vector may be transduced along with the pLentG-KOSM vector.

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

• Reporter Convenience: Includes GFP reporter gene to monitor lentiviral 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 KOSM lentivirus, please see our ViraSafe™ Lentiviral Packaging Systems on page 46.


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

26

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 56.

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

27

Our murine embryonic fibroblast (MEF) feeder cells are useful for the maintenance of hES cells in their undiffer-entiated 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.


28

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

5 x 96 Assays CBA-320-5

• 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


29

StemTAG™ 96-Well Stem Cell Colony Formation Assay


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

5 x 96 Assays CBA-325-5

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

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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 Complete Kit ICC & Colorimetric 2 x 100 Assays CBA-302

StemTAG™ Alkaline Phosphatase Activity Assay Kit Colorimetric 100 Assays CBA-301

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

Alkaline Phosphatase (AP) is a widely used marker for both mouse and human embryonic stem (ES) cells and embryonic germ (EG) cells. The Stem-TAG™ Alkaline Phosphatase Kits provide an efficient system for monitoring cell differentiation or undiffer-entiation using the AP marker. The StemTAG™ Alkaline Phosphatase Complete Kit provides reagents for monitoring AP activity via im-munocytochemistry staining and a quantitative colori-metric activity assay, with sufficient reagents for 100 assays of each method. The ICC and colorimetric 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. 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)

2. 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)

3. 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.

4. 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. 33-36

Adenovirus p. 37-44

Lentivirus (HIV-1, FIV, SIV)

p. 45-51

Retrovirus (MMLV) p. 52-60

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 Yes 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

32

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


VIRAL EXPRESSION AAV Expression

33

Adeno-Associated Virus Kits & Reagents

Adeno-associated virus (AAV) integrates into the host cell genome at a very specific site in one human chromosome. Random insertions are rare, making AAV 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 Expression System

Ordering information for AAV Helper Free Expression Systems, Packaging Systems, Expression Vectors, and Control Plasmids may be found on the following page.

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

• Control Vectors: GFP, Cre or LacZ

• Purification Kits • Quantitation / Titer Kit • Transduction Kits

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

Gene Delivery using the AAV Helper Free System.

Transduction of AAV-GFP using the AAV Helper Free System. GFP expression in 293AD cells 48 hours after transduction (20X).



34


AAV-2 Helper Free Expression System 1 kit VPK-410-SER2

AAV-2 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-SER2

AAV-2 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER2

AAV-2 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER2

AAV-2 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-SER2

AAV-2 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-SER2

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 • pRep-Cap Plasmid • AAV Expression Vector • GFP Control Vector

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 system is ideal if you already have an AAV expression construct.


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

AAV Helper Free 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

AAV Helper Free Control Vectors


pAAV-GFP Control Vector 10 µg AAV-400

pAAV-Cre Control Vector 10 µg AAV-401

pAAV-LacZ Control Vector 10 µg AAV-402

COMING SOON: Helper Free Expression Systems for additional AAV serotypes. Visit

www.cellbiolabs.com for the latest information.



35

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 specific for AAV-2 serotype.

• 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 2 x 10cm dishes 10 Preps VPK-140

ViraBind™ AAV Purification Mega Kit 2 Preps VPK-141

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

Our ViraBind™ AAV Purification Kits are available in two sizes: • The Standard kit can process 1 x 15cm dish or 2 x

10cm dishes per prep • The Mega kit can process 10 x 15cm dishes per

prep

Purification Procedure for the ViraBind™ AAV Purification Kit.

AAV Packaging 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.



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 purified AAV.

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AAV DNA STD (ng)

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AAV DNA STD (ng)

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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

36

*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

37

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

• Premade Recombinant Adenoviruses • Viral Expression Systems • Viral Packaging Cell Line

RAPAd® Adenoviral Expression Systems

*Allows you the flexibility to clone your own promoter along with your gene of interest.

Adenovirus Production using the RAPAd® Adenoviral Expression System.

Adenoviruses are useful for delivering genes into a wide variety of cell types. Traditional recombination methods, however, take several months and require tedious plaque purification. More recent technolo-gies have shortened the time required, but these methods still produce relatively high levels of wild-type (replication-competent) plaques, levels that in-crease even further with serial amplification. The RAPAd® Adenoviral Expression Systems pro-duce recombinant adenovirus with substantial re-duction in wild-type adenovirus, while doing so in a much shorter 2 week time frame. The 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 Virus Production: Virus generated in 2 weeks compared to a few months with traditional methods

• 5 Complete Systems: Choose CMV or RSV as your promoter for gene expression, EF-1 for miRNA expression, or clone your own promoter along with your gene of interest for maximum versatility

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


38


Don’t have time to make your own adenovirus? Are you studying the expression of multiple genes? Rely on our premade recombinant adenoviruses that already contain a gene of interest. 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.

Target Name Catalog Number

Null Control (No gene) ADV-001

β-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. 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. (ADV-002)

3. Takeda, R. et al (2008). Calcineurin is critical for sodium-induced neointimal formation in normotensive and hypertensive rats. Am. J. Physiol. Heart Circ. Physiol. 294:H2871-H2878. (ADV-002)

4. 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-004)

5. Perez-Moreno, M. et al. (2008). Loss of p120 catenin and links to mitotic alterations, inflammation, and skin cancer. PNAS 105:15399-15404. (ADV-004)

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-1α ADV-100

Recent Product Citations 1. 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)

2. Stoletov, K. et al (2008). High resolution imaging of the dy-namic tumor cell-vascular interface in transparent zebrafish. PNAS 104:17406-17411. (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. Ponugoti, B. et al (2007). Functional interaction of HYNF-4 and

PGC-1α in CYP7A1 regulation is inhibited by a key lipogenic activator, SREBP-1c. Mol. Endocrinol. 21:2698-2712. (AD-100)

2. Fang, S. et al (2007). Coordinated recruitment of histone me-thyltransferase G9a and other chromatin modifying enzymes in SHP-mediated regulation of hepatic bile acid metabolism. Mol. Cell Biol. 27:1407-1424. (AD-100)


293AD Cell Line 1 x 106 Cells AD-100



39

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


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

3. 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)

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)


6. 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)

7. 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)

8. Fang, W.B. et al. (2008). Overexpression of EPHA2 receptor destabilizes adherens junctions via a RhoA-dependent mecha-nism. J. Cell Sci. 121:358-368. (ADV-157)

9. Moldobaeva, A. et al. (2008). MIP-2 causes differential activa-tion of RhoA in mouse aortic versus pulmonary artery endothe-lial cells. Microvascular Res. 75:53-58. (ADV-157)

Cell Cycle


p53 ADV-501

p53 (Temperature Sensitive Mutant) ADV-502

p68 RNA Helicase ADV-505



DCC ADV-504

MyoD ADV-508

Myogenin ADV-509


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-1α ADV-210

40


MAP Kinase Signaling



Cdc42 ADV-152



ERK2 ADV-112

ERK2 (Dominant Negative) ADV-113

ERK5 (BMK1) ADV-116

ERK5 (Dominant Negative) ADV-117

Interferon-γ 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

p38α ADV-104

p38α (Dominant Negative) ADV-105

p38β ADV-106

p38β (Dominant Negative) ADV-107

p38γ ADV-108

p38γ (Dominant Negative) ADV-109

p38δ (Dominant Negative) ADV-111

PRAK (Dominant Negative) ADV-141





MKK6 ADV-123

MKK3 ADV-120

See following page for recent citations using these adenoviruses


41



Tyrosine Kinases and PKCs


CSK ADV-405

CSK (Dominant Negative) ADV-406

Fyn ADV-403

Fyn (Dominant Negative) ADV-404

PKC-α (Dominant Negative) ADV-410

PKC-θ (Dominant Negative) ADV-411

PKC-ζ (Dominant Negative) ADV-412

shAkt1 ADV-417

shAkt2 ADV-418

Src ADV-401

Recent Product Citations 1. 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)

2. 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)

3. Naimi, M. et al (2007). Nuclear forkhead box O1 controls and integrates key signaling pathways in hepatocytes. Endocrinol-ogy 148:2424-2434. (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 protease inhibitor ritonavir inhibits cholesterol efflux from human macro-phage-derived foam cells. Am. J. of Pathology 171:304-314. (ADV-113)

8. Black, S. et al (2007). Tissue-specific mechanisms for CCN2/CTGF persistence in fibrotic gingiva. J. Biol. Chem. 282(21):15416-15429. (ADV-115)

9. Vadysirisack, D. et al (2007). MEK signaling modulates sodium iodide symporter at multiple levels and in a paradoxical man-ner. Endocrine-Related Cancer 14:421-432. (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. 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)

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

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

16. 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)

17. Fang, W.B. et al (2008). Overexpression of EPHA2 receptor destabilizes adherens junctions via a RhoA-dependent mecha-nism. J. Cell Sci. 121:358-368. (ADV-157)

18. 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 NFκB Signaling


IκB-α ADV-301

IκB-α S32A (Dominant Negative) ADV-302

IKK-β (Dominant Negative) ADV-303

NOD2 ADV-308

Rel B ADV-304

IKK-β ADV-305

NOD2 (Frame Shift Mutation) ADV-309

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. 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. (ADV-302)

3. 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)

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

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

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



42

ViraBind™ Adenovirus Purification Kits

Purification of Recombinant Ad-β-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 Purification Kit 2.5 x 1012 VP 10 Preps VPK-100

ViraBind™ Adenovirus Purification Mega Kit 2.5 x 1013 VP 1 Prep VPK-101

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

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

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)

Recent Product Citations 1. 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)

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

3. Sukumaran S. et al. (2009). Functional characterization of the human 1-acylglycerol-3-phosphate-O-acyltransferase isoform 10/glycerol-3-phosphate acyltransferase isoform 3. J. Mol. Endocrinol. 42:469-478. (VPK-100)

4. Li, H. et al. (2009). Nuclear translocation of Ad/EYFP-hCAR: a novel tool for screening human CAR activators in human pri-mary hepatocytes. Drug Metab. Dispos. 37:1098-1106. (VPK-100)

5. Guan, M. et al. (2008). Adenovirus-mediated restoration of expression of the tumor suppressor gene DLC1 inhibits the proliferation and tumorigenicity of aggressive, androgen-independent human prostate cancer cell lines: prospects for gene therapy. Cancer Gene Ther. 15:371-381. (VPK-100)

6. Tragoolpua, K.et al. (2008). Generation of functional scFv in-trabody to abate the expression of CD147 surface molecule of 293A cells. BMC Biotechnology 8:5. (VPK-100)

7. Kang, S-H. et al. (2008). Roles of ERK and NFkB in interleukin-8 expression in response to heat shock protein 22 in vascular smooth muscle cells. Korean J. Physiol. Pharmacol. 12:171-176. (VPK-100)

8. Soesanto, Y. et al (2008). Regulation of Akt signaling by OGl-cNAc by euglycemia. Am. J. Physiol. Endocrinol. Metab. 295:E974-E980. (VPK-100)

9. Coleman, A. et al (2008). Cooperative regulation of non-small cell lung carcinoma angiogenic potential by macrophage migra-tion inhibitory factor and its homolog, D-Dopachrome Tautom-erase. J. Immunol. 181:2330-2337. (VPK-100)

10.Tchivilev, I. et al (2008). Identification of a protective role for protein phosphatase 1c-gamma-1 against oxidative stress-induced vascular smooth muscle cell apoptosis. J. Biol. Chem. 283:22193-22205. (VPK-100)

11.Scott, M. et al (2008). ß2-integrin induced p38MAPK activation is a key mediator in the CD14/TLR4/MD2-dependent uptake of LPS by hepatocytes. J. Biol. Chem. 283:29433-29446. (VPK-100)

Selection Guide for ViraBind™ Adenovirus Purification Kits

ViraBind™ Adenovirus Miniprep Kit

ViraBind™ Adenovirus Purification Kit

ViraBind™ Adenovirus Purification Mega Kit

Purification Method Spin column Syringe filter Syringe filter

Purification Time 30 minutes 30 minutes 1-2 hours

Capacity/Prep (Viral Particles) 1 x 1011 VP 2.5 x 1012 VP 2.5 x 1013 VP

Capacity/Prep (Supernatant) One T75 flask

or one 10cm dish Four T75 flasks 50 T75 flasks



43

QuickTiter™ Adenoviral Titer & Quantitation Kits

QuickTiter™ Adenovirus Titer Immunoassay Kit. 293AD cells (p. 40) 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-

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. 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)

2. Kang, S-H. et al. (2008). Roles of ERK and NFkB in interleukin-8 expression in response to heat shock protein 22 in vascular smooth muscle cells. Korean J. Physiol. Pharmacol. 12:171-176. (VPK-106)

3. Peled, M. et al. (2009). Systemic administration of a conditionally replicating adenovirus, targeted to angiogenesis, reduced lung metastases burden in cotton rats. Clin. Cancer Res. 15:1664-1673. (VPK-109)

4. Troidl, K. et al. (2009). Actin-binding Rho activating protein (Abra) is essential for fluid shear stress-induced arteriogenesis. Arterio-scler. Thromb. Vasc. Biol. 29(12):2093-2101. (VPK-109)

5. Soesanto, Y. et al (2008). Regulation of Akt signaling by OGl-cNAc by euglycemia. Am. J. Physiol. Endocrinol. Metab. 10.1152/ajpendo.90366.2008. (VPK-109)

6. Chang, C-T. et al (2008). TGF-ß1 decreases epithelial sodium channel functionality in renal collecting duct cells using a Smad4-dependent pathway. Nephrol. Dial. Transplant. 23:1126-1134. (VPK-109)

7. Hoashi, T. et al. (2009). The secreted form of a melanocyte mem-brane-bound glycoprotein (Pmel17/gp100) is released by ectodo-main shedding. FASEB J. 10.1063/fj.09-140921. (VPK-110)

8. Tudhope, S. et al (2007). The role of IκB kinase 2, but not activa-tion of NF-κB, in the release of CXCR3 ligands from IFNγ-stimulated human bronchial epithelial cells. J.Immunol. 179:6237-6245. (VPK-110)

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



44

Rapid Replication Competent Adenovirus (RCA) Assay Kit


Rapid RCA Assay Kit ICC Staining 30 Assays VPK-111

This kit uses the assay principles of the QuickTiter™ Adenovirus Titer Immunoassay Kit (see page 41), 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.

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Recent Product Citations 1. Ackerman, W. et al (2008). Nuclear Factor-kappa B regu-

lates inducible prostaglandin E synthase expression in hu-man amnion mesenchymal cells. Biol. Reprod. 78:68-76. (AD-200, AD-201)

2. Monick, M. et al (2008). Constitutive ERK MAPK activity regulates macrophage ATP production and mitochondrial integrity. J. Immunol. 180:7485-7496. (AD-200)

*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.


45

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 page 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: Nine cloning vectors to choose from; compatible with any 2nd or 3rd generation packaging system


46


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


Ecotropic 1 Kit VPK-218-ECO


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


ViraSafe™ miRNA Lentiviral Expression System Ecotropic 1 Kit VPK-220-ECO


ViraSafe™ Lentiviral Bicistronic Expression System (GFP)

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; VSVG for any species.

ViraSafe™ Lentiviral Packaging Systems

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


ViraSafe™ Lentiviral Packaging System Colorimetric 96 Assays VPK-205

5 x 96 Assays VPK-206

293LTV Lentiviral Cell Line


293LTV Cell Line 1 x 106 Cells LTV-100

Recent Product Citations Shimamura, T. et al (2008). Hsp90 inhibition suppresses mutant EGFR-T790M signaling and overcomes kinase inhibitor resistance. Cancer Res. 68:5827-5838. (LTV-100)


47


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 miRNA Lentiviral Expression Vector 10 µg VPK-220

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

These lentiviral expression vectors contain multiple cloning sites to accommodate your gene of interest. Each vector may be used with any 2nd or 3rd gen-eration lentiviral packaging system including our ViraSafe™ Lentiviral Packaging Systems. Each vector can accommodate large inserts of up to at least 7 kb.*

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


GFP Lentivirus Control 200 µL LTV-300

RFP Lentivirus Control 200 µL LTV-301

ß-Galactosidase Lentivirus Control 500 µL LTV-302

Concentration

1 x 106 TU/mL

1 x 106 TU/mL

1 x 105 TU/mL

*Our Universal (Promoterless) vector can accommodate inserts of up to 10 kb total (including promoter).

Map of pSMPUW-IRES-GFP Vector.

These plasmids allow you to package your own reporter lentivirus.


These controls provide the convenience of a pre-made virus packaged with a reporter gene. Each virus is provided at a predetermined concentration.

Lentiviral Trans-duction of 293 Cells. Left: GFP Lentivirus Control Right: ß-Gal Len-tivirus Control.

Assay Principle for the QuickTiter™ Lentivirus Titer Kit. Upon transfection of lentiviral packaging plasmids into 293T cells, lenti-virus particles are packaged with p24 protein. However, additional free p24 protein not associated with virus is present in viral super-natant. A traditional p24 ELISA will detect both sources of p24 which overestimates viral titer. The QuickTiter™ Lentivirus Titer Kit uses proprietary technology to pull the virus out of solution prior to quantitation, providing a more accurate measurement of viral titer.

48


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 kits provide a quick, convenient way to quantify the concentration of your lentivirus. Individual kits are available to measure either HIV-1 or FIV concentra-tions. 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™ 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 or FIV 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

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49


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

5 x 96 Assays VPK-107-5

QuickTiter™ HIV Lentivirus Quantitation Kit (HIV-1 p24 ELISA) Colorimetric 96 Assays VPK-108-HIV-P24

5 x 96 Assays VPK-108-HIV-P24-5

QuickTiter™ FIV Lentivirus Quantitation Kit (FIV p24 ELISA) 96 Assays VPK-108-FIV-P24

5 x 96 Assays VPK-108-FIV-P24-5

QuickTiter™ Lentivirus Quantitation Kit Fluorometric 20 Assays VPK-112

Colorimetric

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. Tiedemann, R.E. et al. (2010). Kinome-wide RNAi studies in human

multiple myeloma identify vulnerable kinase targets, including a lym-phoid restricted kinase, GRK6. Blood 115:1594-1604. (VPK-107)

2. Lin, R-J. et al. (2009). Distinct antiviral roles for human 2’5’-oligoadenylate synthetase family members against dengue virus infec-tion. J. Immunol. 183:8035-8043. (VPK-107)

3. Sukumaran S. et al. (2009). Functional characterization of the human 1-acylglycerol-3-phosphate-O-acyltransferase isoform 10/glycerol-3-phosphate acyltransferase isoform 3. J. Mol. Endocrinol. 42:469-478. (VPK-107)

4. Joshi, M.B. et al. (2009). Extracellular cadherin repeat domains EC1 and EC5 of T-cadherin are essential for its ability to stimulate angiogenic behavior of endothelial cells. FASEB J. 10.1096/fj.09/133611. (VPK-107)

5. Keck, Z.-y. et al. (2009). Mutations in HCV E2 located outside the CD81 binding sites lead to escape from broadly neutralizing antibodies but compromise virus infectivity. J. Virol. 83:6149-6160. (VPK-107)

6. Escamilla-Hernandez, R. et al. (2008). Constitutively active protein kinase A qualitatively mimics the effects of follicle-stimulating hormone on granulosa cell differentiation. Mol. Endocrin. 22(8):1842-1852. (VPK-107)

7. Dyer, K. et al. (2008). Functionally competent eosinophils differentiated ex vivo in high purity from normal mouse bone marrow. J. Immunol. 181:4004-4009. (VPK-107)

8. Matsumae, H. et al. (2008). CCN1 knockdown suppresses neointimal hyperplasia in a rat artery balloon injury model. Arterioscler Thromb Vasc Biol. 28(6):1077-83. (VPK-108-FIV-P24)

9. Brittain, J.M. et al. (2009). An atypical role for collapsing response me-diator protein (CRMP-2) in neurotransmitter release via interaction with presynaptic voltage-gated calcium channels. J. Biol. Chem. 284:31375-31390. (VPK-108-HIV-P24)

10. Agrawal-Gamse, C. et al. (2009). Adaptive mutations in a human immu-nodeficiency virus type 1 envelope protein with a truncated V3 loop restore function by improving interactions with CD4. J. Virol. 83:11005-11015. (VPK-108-HIV-P24)

11. Kaletsky, R. et al. (2009). Tetherin-mediated restriction of filovirus bud-ding is antagonized by the Ebola glycoprotein. PNAS 106:2886-2891. (VPK-108-HIV-P24)

12. Lambert, M.P. et al. (2009). Platelet factor 4 regulates megakaryopoi-esis through low density lipoprotein receptor related protein 1 (LRP1) on megakaryocytes. Blood 114:2290-2298. (VPK-108-HIV-P24)

13. Malur, A. et al. (2009). Deletion of PPAR-gamma in alveolar macro-phages is associated with Th-1 pulmonary inflammatory response. J. Immunol. 182:5816-5822. (VPK-108-HIV-P24)

14. Shu, L. and P.J. Houghton. (2009). The mTORC2 complex regulates terminal differentiation of C2C12 myoblasts. Mol. Cell Biol. 29:4691-4700. (VPK-108-HIV-P24)

15. Krishnamachary, B. et al. (2009). Noninvasive detection of lentiviral-mediated choline kinase targeting in a human breast cancer xenograft. Cancer Res. 69:3464-3471. (VPK-108-HIV-P24)

16. Wilson, A.A. et al. (2008). Sustained expression of alpha-1 antitrypsin after transplantation of manipulated hematopoietic stem cells. Am. J. Respir. Cell Mol. Biol. 39:133-141. (VPK-108-HIV-P24)

17. Niwano, K. et al. (2008). Lentiviral vector-mediated SERCA2 gene trans-fer protects against heart failure and left ventricular remodeling after myocardial infarction in rats. Mol. Ther. 16:1026-1032. (VPK-112)



50

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



51

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 Vira-Ductin™ 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®.

The efficiency of lentivirus transduction is typically low. Additives such as Polybrene® can boost trans-duction efficiencies, but even then only a small frac-tion of lentiviral 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®. The ViraDuctin™ system is ideal for primary cells and stem cells.

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

• More Robust: Useful for transduction of non-permissive 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.


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VIRAL EXPRESSION Retroviral Expression

52

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:

• Retroviral Packaging Cell Lines • Concentration / Purification Kits • Quantitation / Titer Kits • Transduction Reagents

• Gene-Specific Retroviral Vectors • Retroviral Cloning & Expression

Vectors • Retroviral Expression Systems


c-Abl RTV-402

c-Abl-TM RTV-403

c-Abl (1-565) RTV-404

c-Abl (1-958) RTV-405

p53 RTV-401

Vector Backbone

pBABEpuro

pBABEpuro

pBABEpuro

pBABEpuro

pBABEpuro

Cell Cycle

Reporter Genes

If you are studying multiple genes, you might appreci-ate the convenience of our gene-specific retroviral constructs. These vectors are based on backbones derived from the Moloney murine leukemia virus (MMLV). Each vector contains a gene of interest from a variety of pathways and disciplines. Vectors containing GFP or iPS stem cell factors are supplied as 10 µg of plasmid in TE buffer. All other vectors are supplied as 100 µL of bacterial glycerol stock.

Gene-Specific Recombinant Retroviral Vectors


GFP RTV-002

GFP RTV-051

Vector Backbone

pBABE

pMCs

GFP pMX RTV-050

GFP pMYs RTV-052

Generic Map of pBABEhygro Retroviral Expression Vector. 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)

Autophagy


GFP-LC3 pMXs RTV-801

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



53

Target Name Vector Backbone Mutation State Catalog Number

Cdc42 pBABEhygro L61 RTV-203

pBABEpuro N/A RTV-220

pWZLhygro Q61 RTV-221

myr-Rac1 pBABEpuro N/A RTV-201

Rac1 pBABEhygro V12 RTV-202

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

K-Ras

N-Ras pBABEpuro K61 RTV-222

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-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


Human iPS Genes Mouse iPS Genes

Proteases

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


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

p38α pBABEhygro Dominant Negative RTV-105

p38β pBABEhygro Dominant Negative RTV-106

p38γ pBABEhygro Dominant Negative RTV-107

p38δ pBABEhygro Dominant Negative RTV-108

PI3K p110α-CAAX pWZLneo Constitutively Active RTV-124

pBABEpuro Constitutively Active RTV-122


Raf1-CAAX pWZLneo Constitutively Active RTV-113

PRAK

MAP Kinase Signaling

54



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

Transcription Regulation

Gene-Specific Recombinant Retroviral Vectors, continued


Stat5A(1*6) pMXs RTV-331

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



55

Retroviral Cloning & Expression Vectors

Our Retroviral Expression Vectors contain multiple cloning sites (MCS) to allow easy introduction of your gene of interest. The vectors are based on backbones derived from Moloney murine leukemia virus (MMLV). The backbone is either the traditional pBABE system or the newer pMXs/pMYs/pMZs system, which has been shown to be useful in induced pluripotent stem cell (iPS) studies. Specifically pMYs vectors are opti-mal for use with hematopoietic 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 Catalog Number

pMXs-IRES-Puro RTV-014

pMXs-Neo RTV-011

pMXs-Puro RTV-012

pMYs RTV-020

pMYs-IRES-GFP RTV-021

pMYs-IRES-Neo RTV-023

pMYs-IRES-Puro RTV-022

pMYs-Puro RTV-024

pMZs RTV-030

pBABEhygro RTV-001-HYGRO

pBABEneo RTV-001-NEO

pBABEpuro RTV-001-PURO

pMCs-IRES-GFP RTV-040

pMCs-Puro RTV-041

pMXs RTV-010

pMXs-IRES-Blasticidin RTV-016

pMXs-IRES-GFP RTV-013

pMXs-IRES-Neo RTV-015

pMXs-miR-GFP/Puro RTV-017

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.

Our 293RTV cells are derived from the 293 parental cell line, but are selected for firmer attachment to cul-ture plates, faster growth and higher yields of retrovi-rus produced.


pCMV-10A1 Envelope Vector 100 µL RV-114

pCMV-Ampho Envelope Vector 100 µL RV-113

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

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

pCMV-Eco Envelope Vector 100 µL RV-112

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


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

56


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. +++

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

High Viral Yields with Plat-E Cells. ProB Ba/F3 cells were infected with GFP retrovirus supernatant pro-duced in Plat-E cells after transfec-tion with pMX-GFP vector.

Suitability of Platinum Retroviral Packaging Cell Lines by Host Species.

Platinum Retroviral Expression Systems

• 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

Retrovirus Production Using the Platinum Retroviral Expression Systems.

Retroviral vectors are useful for delivering genes of interest into a host cell where integration into the ge-nome is desired. However, traditional retroviral ex-pression technologies usually result in low viral titers, making gene expression studies difficult. This is mainly due to poor expression of retroviral structure proteins (e.g. gag, pol, and env) in the 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. 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.

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.

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



57


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 Expression System, Ecotropic

1 kit VPK-300

Platinum Retroviral Expression System, Amphotropic

1 kit VPK-301

Platinum Retroviral Expression System, Pantropic

1 kit VPK-302

Platinum ES/EC Retroviral Expression System, Ecotropic

1 kit VPK-303

Platinum ES/EC Retroviral Expression System, Amphotropic

1 kit VPK-304

Platinum ES/EC Retroviral Expression System, Pantropic

1 kit VPK-305

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

Packaging Cell

Plat-E

Plat-A

Plat-GP

Plat-E

Plat-A

Plat-GP

Plat-E

Plat-A

Plat-GP

Expression Vector

pMXs-Puro

pMXs-Puro

pMXs-Puro

pMCs-Puro

pMCs-Puro

pMCs-Puro

pMYs-Puro

pMYs-Puro

pMYs-Puro

Platinum Retroviral Expression Systems, continued

Platinum Retroviral Expression Systems contain everything you need to generate your recombinant retrovirus: packaging cell line, expression vector, and GFP control vector. Our pantropic systems also contain a VSVG en-velope vector. Simply clone your gene of interest into the expression vector. These complete systems offer the greatest value, but you may also order the Platinum Packaging Cell Lines and Retroviral Expression Vectors separately on this page and the next.

Platinum Retroviral Packaging Cell Lines

Recent Product Citations 1. Lee, J-H. et al. (2010). Epigallocatechin-3-gallate inhibits osteoclasto-

genesis by downregulating c-fos expression and suppressing the nu-clear factor k-B signal. Mol. Pharmacol. 77:17-25. (RV-101)

2. Rice, K.L. et al. (2009). Comprehensive genetic screens identify a role for PLZR-RAR-alpha as a positive regulator of cell proliferation via direct regulation of c-myc. Blood 114:5499-5511. (RV-101)

3. Cui, G. et al. (2009). Berberine differentially modulates the activities of ERK, p38 MAPK, and JNK to suppress Th17 and Th1 T cell differentia-tion in Type 1 diabetic mice. J. Biol. Chem. 284:28420-28429. (RV-101)

4. Huse, J.T. et al. (2009). The PTEN-regulating microRNA mir-26a is amplified in high-grade glioma and facilitates gliomagenesis in vivo. Genes & Dev. 23:1327-1337. (RV-101)

5. Hinrichs, C.S. et al. (2009). Adoptively transferred effector cells derived from naive rather than central memory CD8+ cells mediate superior antitumor immunity. PNAS 106:17469-17474. (RV-101)

6. Wu, Y. et al. (2009). Improved coinfection with amphotropic pseudo-typed retroviral vectors. J. Biochem. Biotechnol. 10.1155/2009/901079. (RV-102)

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-GP cells require co-transfection with VSVG envelope vector (available separately).



58

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.



59

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 content of purified virus or unpurified viral super-natant. 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.


60

ViraDuctin™ Retrovirus Transduction Kit


40 Transductions RV-200

200 Transductions RV-201 ViraDuctin™ Retrovirus Transduction Kit

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

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.

The efficiency of retrovirus transduction can be low compared to other viruses. The rate at which retroviral vec-tors bind to cells is controlled mostly by diffusion. Additionally, the presence of transduction inhibitors such as proteoglycans and glycosaminoglycans in retroviral supernatants can lead to poor gene transfer. Additives such as Polybrene® can boost transduction efficiencies, but they do not eliminate these transduction inhibitors.

Recent Product Citation Miyoshi, N. et al. (2010). Defined factors induce reprogramming of gastroin-testinal cancer cells. PNAS 107:40-45.


QuickTiter™ HBV and HCV Core Antigen ELISA Kits

These kits specifically quantify the core antigens of Hepatitis B and Hepatitis C viruses, respectively. 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.

• 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 Colorimetric 96 Wells VPK-150

QuickTiter™ HCV Core Antigen ELISA Kit Colorimetric 96 Wells VPK-151

0

0.5

1

1.5

2

2.5

3

0 20 40 60 80 100 120

HBVcAg p24 (ng/mL)

OD

45

0n

m

Standard Curve Generated with the QuickTiter HBV Core Antigen ELISA Kit.

62

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


miRNASelect™ Expression Vectors contain full miRNA precursor se-quences with constitutive promoter. • Vectors with human (hsa) se-

quences contain a puromycin selection marker

• Vectors with mouse (mmu)sequences contain a GFP-puromycin fusion to allow se-lection by either marker


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


hsa-mir-16-1 MIR-16-1



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-26a-1 MIR-26A-1




hsa-mir-26b MIR-26B

hsa-mir-27a MIR-27A

hsa-mir-27b MIR-27B


hsa-mir-28 MIR-28


hsa-mir-29a MIR-29A





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-29c MIR-29C

mmu-mir-29c MMU-MIR-29C

hsa-mir-30a MIR-30A


Don’t see your miRNA of interest? Clone your own se-quence into one of our Expression Vectors—see page 68.


63


miRNASelect™ Precursor Clone Collection (Expression Vectors), cont.



mmu-mir-30d MMU-MIR-30D

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-100 MIR-100


hsa-mir-30d MIR-30D

hsa-mir-30e MIR-30E

mmu-mir-30e MMU-MIR-30E






hsa-mir-96 MIR-96


hsa-mir-98 MIR-98



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-129-2 MIR-129-2






hsa-mir-132 MIR-132
















hsa-mir-142 MIR-142

hsa-mir-143 MIR-143





hsa-mir-136 MIR-136

hsa-mir-137 MIR-137

hsa-mir-138-2 MIR-138-2


hsa-mir-138-1 MIR-138-1


hsa-mir-139 MIR-139

hsa-mir-140 MIR-140




hsa-mir-145 MIR-145


hsa-mir-147 MIR-147




hsa-mir-151 MIR-151


hsa-mir-152 MIR-152


hsa-mir-150 MIR-150





64




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-190 MIR-190







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-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








hsa-mir-298 MIR-298











65








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











hsa-mir-491 MIR-491










66

Product Name Catalog Number

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











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-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-600 MIR-600

hsa-mir-601 MIR-601

hsa-mir-602 MIR-602

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-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


hsa-mir-542 MIR-542


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




67























hsa-mir-708 MIR-708













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-876 MIR-876


hsa-mir-877 MIR-877






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-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-933 MIR-933

hsa-mir-934 MIR-934

hsa-mir-935 MIR-935

hsa-mir-936 MIR-936

hsa-mir-921 MIR-921

hsa-mir-922 MIR-922

hsa-mir-923 MIR-923

hsa-mir-924 MIR-924























68

Expression, Control, Reporter Vectors

miRNASelect™ Expression and Control Vectors

MICRORNA ANALYSIS


100 µL MIR-EXP-GP-C miRNASelect™ pEGP-mir Cloning and Expression Vector

miRNASelect™ pEP-mir Cloning and Expression Vector 100 µL MIR-EXP-C

miRNASelect™ pEGP-mir Null Control Vector 100 µL MIR-NULL-GP

miRNASelect™ pEP-mir Null Control Vector 100 µL MIR-NULL

Our miRNASelect™ Mammalian Expression Vectors provide an easy, efficient method to clone a miRNA precursor from any species. The desired miRNA se-quence is cloned into a human ß-globin intron con-tained within the vector. Two vector formats are avail-able: • 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. Null control vectors are also sold separately for use with vectors from our miRNASelect™ Precursor Clone Collection.

miRNASelect™ Functional Analysis Reporter System


1 Kit MIR-GFP miRNASelect™ pMIR-GFP Reporter System

Assay Principle. If miRNA is pre-sent and binds to the 3’ UTR, trans-lation of the GFP gene is repressed, resulting in loss of fluorescence.

The miRNASelect™ Functional Analysis Reporter System provides a simple method for the evalua-tion of potential targets of miRNA. Binding of miRNA sequences to their suspected targets re-sults in repression of translation. 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 immedi-ately downstream of the GFP gene. If the miRNA is present and binds to the target sequence, trans-lation is repressed and no green fluorescence ap-pears. If the miRNA does not bind the target se-quence, GFP translation occurs normally and green fluorescence may be seen.


ViraSafe™ miRNA Lentivirus Expression System

• Safer: 80-90% less sequence homology than 3rd-generation lentiviral expression systems; ecotropic system provides even more safety*

• High Titer: Incorporates elements that provide titers comparable to 3rd-generation systems

• Flexible: Vectors provided separately for in-creased safety and to allow optimization of vec-tor ratios

Our ViraSafe™ Lentiviral Expression Systems pro-vide a much safer method even compared to third-generation lentivirus systems. Sequence homology has been reduced an additional 80-90%, thereby sub-stantially reducing the risk of replication-competent lentivirus. Additionally, each vector is provided sepa-rately to allow for optimization of vector ratios. The ViraSafe™ miRNA Lentivirus Expression System is specifically designed to express microRNA in your target cell.

For more information on our complete se-lection of ViraSafe™ Lentivirus Expression Systems for gene expression studies, please see p. 45-46.

MICRORNA ANALYSIS Viral Expression Systems


ViraSafe™ miRNA Lentiviral Expression System Ecotropic 1 Kit VPK-220-ECO


pSMPUW-miR-GFP-Puro Lentiviral Expression Vector N/A 10 µg VPK-220

69

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

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 systems use a backbone vector from which the 5’ ITR, packaging signal and E1 sequences have been removed. Additionally, serial amplification of the recombinant adenovirus does not increase the level of replica-tion-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 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 Expression Systems for gene expression studies, please see page 37.

The RAPAd® miRNA Adenoviral Expression System is specifically designed to deliver miRNA sequences into your target cell.


Viral Vectors, Knockdown Enhancer

miRNA Retroviral Expression Vector

MICRORNA ANALYSIS

70

RNAi Enhancer Reagent for miRNA and siRNA


20 reactions RNAI-200

100 reactions RNAI-201 RNAi Enhancer Reagent

RNA interference can occur in the presence of either siRNA or the mature form of miRNA. The RNAi Enhancer Reagent increases the level of interference in the presence of siRNA. It also increases the rate of processing of pre-miRNA into mature miRNA.


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 57.


OXIDATIVE & CELLULAR STRESS Oxidative Stress Overview

Measuring Oxidative Stress

72

Oxidative Stress Assays by Sample Type

Marker or Type of Damage

Sample Type

Cells Tissues Blood Urine Other

Oxidative Protein Damage

(p. 73-75)

Protein carbonyl content (PCC) X X X

3-Nitrotyrosine X X X

Advanced Glycation End Products (AGE) X X X

Advanced Oxidation Protein Products(AOPP) X X X

Lipid Peroxidation (p. 76-77)

TBARS X X X X

4-Hydroxynonenal (4-HNE) X X X

Malondialdehyde (MDA) X X X X

8-iso-Prostaglandin F2α (8-Isoprostane) X X X X

DNA / RNA Damage (p. 78-80)

8-hydroxyguanosine (8-OHG) X X X X Cerebrospinal fluid

8-hydroxydeoxyguanosine (8-OHdG) X X X X

Abasic (AP) sites X X

Double-strand DNA breaks X

Comet Assay (various types of DNA damage) X

Antioxidants & Antioxidant

Capacity (p. 81-82)

Superoxide Dismutase X X X X

Catalase X X X

Oxygen Radical Antioxidant Capacity (ORAC) X X X Food

samples

Hydroxyl Radical Antioxidant Capacity (HORAC) X X X Food

samples

Reactive Oxygen Species

(p. 83-84)

DCF (universal marker) X

Hydrogen Peroxide X X X X

Oxidative stress may be measured using one of three primary methods. We offer sensitive, user-friendly assays for each method:

Use the following table to determine the best oxidative stress assays for your samples.

• Measure the reactive oxygen species (ROS) directly • Measure the presence of antioxidants • Measure the resulting damage to proteins, lipids, DNA or RNA (most reliable)


OXIDATIVE & CELLULAR STRESS Protein Oxidation Assays

73

Assays for Oxidative 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 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


Nitrotyrosine ELISA Kit Colorimetric 96 Wells STA-305

5 x 96 Wells STA-305-5

Nitrotyrosine Immunoblot Kit Immunoblot/ECL 10 Blots STA-303

Protein Tyrosine Nitration Control (Nitrotyrosine-BSA) Immunoblot/ECL 10 µg STA-304

Nitric oxide influences a variety of biological processes including cell proliferation, apoptosis, neurotoxicity and extracellular matrix remodeling. Nitric oxide reacts with superoxide to form peroxynitrite, which in turn nitrates tyrosine residues in proteins. Nitrotyrosine therefore serves as a marker for peroxynitrite action in a variety of disease states and in conditions of cellular damage.

Formation of 3-Nitrotyrosine During Oxidative Stress.

Protein Nitration by Tetranitromethane using the OxiSelect™ Nitrotyrosine ELISA Kit. STO (MEF) cells were lysed and nitrated with tetranitromethane. The protein 3-nitrotyrosine levels were measured as described in the assay protocol.

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Recent Product Citations 1. Drel, V.R. et al. (2009). Poly(adenosine 5'-diphosphate-ribose)

polymerase inhibition counteracts multiple manifestations of experimental type 1 diabetic nephropathy. Endocrinology 150:5273-5283. (STA-305)

2. Cheah, F.-C. et al. (2009). Airway inflammatory cell responses to intra-amniotic lipopolysaccharide in a sheep model of chorioamnionitis. Am. J. Physiol. Lung Cell Mol. Physiol. 296:L384-L393. (STA-305)

3. Li, X. et al. (2008). Lipoamide protects retinal pigment epithelial cells from oxidative stress and mitochondrial dysfunction. Free Radic. Biol. Med. 44(7):1465-1474. (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.

74

OXIDATIVE & CELLULAR STRESS Protein Oxidation Assays

OxiSelect™ Protein Carbonyl Assay Kits


OxiSelect™ Protein Carbonyl ELISA Kit 96 Wells 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

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).

Standard Curve Generated with the OxiSelect™ Protein Car-bonyl ELISA Kit (STA-310).

Protein Carbonyl Immunoblot Kit • No Molecular Weight Shift: DNPH Derivatization

after immunoblotting allows direct comparison of oxidized and non-oxidized protein fingerprints

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Protein Carbonyl (nmol/mg)

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450

nm

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 pro-vide rapid, efficient methods for detection of protein carbonyls. Three assay formats are available: im-munoblot, ELISA or spectrophotometer. All formats are suitable for use with purified protein, plasma, serum, or cell lysate samples.

Recent Product Citations 1. Jia, L. et al (2007). Acrolein, a toxicant in cigarette smoke,

causes oxidative damage and mitochondrial dysfunction in RPE cells: protection by (R)-alpha-lipoic acid. Invest. Ophthal-mol. Vis. Sci. 48:339-348. (STA-308)

2. Liu, Z. et al. (2007). Hydroxytyrosol protects retinal pigment epithelial cells from acrolein-induced oxidative stress and mito-chondrial dysfunction. J. Neurochem. 103:2690-2700. (STA-308)

3. Maity, P. et al. (2008). Indomethacin, a non-steroidal anti-inflammatory drug, develops gastropathy by inducing reactive oxygen species-mediated mitochondrial pathology and associ-ated apoptosis in gastric mucosa: A novel role of mitochondrial aconitase oxidation. J. Biol. Chem. 284:3058-3068. (STA-308, STA-310)

4. Kang, K.A. et al. (2010). KIOM-4 protects against oxidative stress-induced mitochondrial damage in pancreatic ß-cells via its antioxidant effects. Evid. Based Complement. Altern. Med. 10.1093/ecam/neq007. (STA-310)

5. Cho, H-Y. et al. (2009). Antiviral activity of Nrf2 in a murine model of respiratory syncytial virus (RSV) disease. Am. J. Res-pir. Crit. Care Med. 179:138-150. (STA-310)

6. Neretti, N. et al. (2009). Long lived Indy induces reduced mito-chondrial reactive oxygen species production and oxidative damage. PNAS 106:2277-2282. (STA-310)

7. Paromov, V. et al. (2008). The influence of N-acetyl-L-cysteine on oxidative stress and nitric oxide synthesis in stimulated macrophages treated with a mustard gas analogue. BMC Cell Biology 9:33. (STA-310)

8. Kim, S-H. et al. (2008). Protective effect of esculetin against oxidative stress-induced cell damage via scavenging reactive oxygen species. Acta Pharmacol. Sinica 29(11):1319-1326. (STA-310)

9. Lowry, M. et al (2007). Lung lining fluid glutathione attenuates IL-13 induced asthma. Am. J. Respir. Cell Mol. Biol. 38(5):509-516. (STA-310)


75

OXIDATIVE & CELLULAR STRESS

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 • Convenient: Quantify on a standard microplate

reader • Versatile: Suitable for use with cell lysates, tissue

homogenates, or plasma

Protein Oxidation Assays

OxiSelect™ Advanced Glycation End Product (AGE) ELISA Kits

Advanced glycation end products are formed during the Maillard reaction where reducing carbohydrates react with lysine side chains and N-terminal amino groups of various macromolecules, particularly pro-teins. 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™ AGE ELISA Kit is designed for the rapid detection of generic advanced glycation end product protein adducts. For more specific detection of CML, one of the most common AGE species, choose the OxiSelect™ CML ELISA Kit.

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AGE-BSA (µg/mL)

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m

• Sensitive: Detect AGE formation as low as 250 ng/mL or CML as low as 5 ng/mL

• Versatile: Compatible with cell lysates, blood samples, or purified proteins

Standard Curve Generated with AGE-BSA Provided in the OxiSelect™ AGE ELISA Kit.


OxiSelect™ Advanced Glycation End Product (AGE) ELISA Kit Colorimetric 96 Wells STA-317

OxiSelect™ Nε-(Carboxymethyl) Lysine (CML) ELISA Kit Colorimetric 96 Wells STA-316

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Chloramine Standard Curve Generated with the OxiSelect™ AOPP Assay Kit.

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Untreated Human Serum Albumin and AOPP-HSA Positive Control Tested with the OxiSelect™ AOPP Assay Kit.


Assays 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 Peroxidation Assays allow you to quickly and easily quantify the most common markers and by-products of lipid peroxidation.

OxiSelect™ HNE-His Adduct ELISA Kit


OxiSelect™ HNE-His Adduct ELISA Kit Colorimetric 96 Wells STA-334

HNE-BSA Control N/A 100 µg STA-335

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HNE (4-hydroxynonenal) is a well-known by-product of lipid peroxidation and is widely accepted as a sta-ble marker for oxidative stress. Our OxiSelect™ HNE-His Adduct ELISA Kit pro-vides a simple, user-friendly way to assess HNE-His adduct formation in a variety of sample types.

• Fast: Obtain results in about 4 hours • Versatile: Suitable for use with cell lysates, se-

rum, or plasma samples

OxiSelect™ 8-iso-Prostaglandin F2α ELISA Kit (8-isoprostane)


OxiSelect™ 8-iso-Prostaglandin F2α ELISA Kit Colorimetric 96 Wells STA-337

8-iso-Prostaglandin F2α is produced in membrane phospholipids and has been implicated in athero-genesis, rheumatoid arthritis and carcinogenesis. The OxiSelect™ 8-iso-Prostaglandin F2α ELISA Kit provides rapid detection of 8-iso-PGF2α.

• Sensitive: Detect as little as 50 pg/mL • Fast: Obtain results in less than 3 hours • Versatile: Suitable for use with urine, plasma, se-

rum or cell lysates

76

HNE-BSA ELISA Standard Curve.

Human Urine Tested with the OxiSelect™ 8-isoPGF2α ELISA.

1.5

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OXIDATIVE & CELLULAR STRESS Lipid Peroxidation Assays

Recent Product Citations 1. Mukhopadhyay, P. et al. (2010). CB1 cannabinoid receptors

promote oxidative stress and cell death in murine models of doxorubicin-induced cardiomyopathy in human cardiomyo-cytes. Cardiovasc. Res. 85:773-784. (STA-334)

2. Maki, R.A. et al. (2009). Aberrant expression of myeloperoxi-dase in astrocytes promotes phospholipid oxidation and mem-ory deficits in a mouse model of Alzheimer's disease. J. Biol. Chem. 284:3158-3169. (STA-334)


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; the TBARS assay measures the formation of this adduct. Our OxiSelect™ TBARS Assay Kit provides a more user-friendly protocol for quantitation of the MDA-TBA adduct compared to other commercial assays.


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: No marbles or glass tubes required

as with other TBARS assays • Versatile: Detect with either standard ELISA or

fluorometric plate reader

OxiSelect™ MDA (Malondialdehyde) Assays


OxiSelect™ MDA Immunoblot Kit Immunoblot 10 Blots STA-331

MDA-BSA Control N/A 100 µg STA-333

OxiSelect™ MDA ELISA Kit Colorimetric 96 Wells STA-332

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. Our OxiSelect™ MDA Immunoblot Kit provides a more direct method to quantify MDA formation than the TBARS Assay. The OxiSelect™ MDA ELISA Kit offers a higher throughput format to accommodate a larger number of samples, and can detect 2 pmol/mg.

77

MDA-TBA Standard Curve Using a Standard Plate Reader.

Immunoblot of MDA-BSA Control. MDA-BSA Im-munoblot Control was first electroblotted onto a nitro-cellulose membrane. MDA was detected by im-munoblotting with anti-MDA antibody as described in the assay protocol.

Colorimetric Assay

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D 5

40

nm

OXIDATIVE & CELLULAR STRESS Lipid Peroxidation Assays

Recent Product Citation Barabutis, N. et al. (2008). Antioxidant activity of growth hormone-releasing hormone antagonists in LNCaP human prostate cancer line. PNAS 105:20470-20745. (STA-331)

Recent Product Citations 1. Song, Y.R. et al. (2010). Activation of hypoxia-inducible factor

attenuates renal injury in rat remnant kidney. Nephrol. Dial. Transplant. 25:77-85. (STA-330)

2. Fomenko, D.E. et al. (2009). Methionine-R-sulfoxide reductase 1 (MsrB1) knockout mice: Roles of MsrB1 in redox regulation and identification of a novel selenoprotein form. J. Biol. Chem. 284:5986-5993. (STA-330)

3. Fujita, K. et al. (2008). Effectiveness of antiplatelet drugs against experimental non-alcoholic fatty liver disease. Gut 57:1583-1591. (STA-330)


78

Assays for DNA & RNA Damage

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 / RNA Damage ELISA Kits (8-OHdG or 8-OHG Quantitation) Among numerous 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. The OxiSelect™ Oxida-tive RNA Damage ELISA Kit uses the same assay principle, but is useful for measuring 8-OHG forma-tion, the primary marker for RNA oxidation.

• Highly Sensitive: Detect as little as 100 pg/mL of 8-OHdG or 300 pg/mL of 8-OHG

• Versatile: Suitable for use with urine, serum, cerebrospinal fluid, and cell or tissue samples


OxiSelect™ Oxidative DNA Damage ELISA Kit (8-OHdG Quantitation) 96 Wells STA-320


OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation) Colorimetric 96 Wells STA-325


Colorimetric

Standard Curve Generated with the OxiSelect™ Oxidative DNA Damage ELISA Kit.

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8-OHdG (ng/mL)

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m

Recent Product Citations 1. Hasegawa, T. et al. (2009). Suppression of nitrosative and oxida-

tive stress to reduce cardiac allograft vasculopathy. Am.J. Physiol. Hear Circ. Physiol. 296:H1007-H1016. (STA-320)

2. Pialoux, V. et al. (2009). Effects of exposure to intermittent hy-poxia on oxidative stress and acute hypoxic ventilatory response in humans. Am. J. Respir. Crit. Care Med. 180:1002-1009. (STA-320)

3. Ksiazek, K. et al. (2008). Vulnerability to oxidative stress and differen patterns of senescence in human peritoneal mesothelial cell strains. Am J. Physiol. Regulatory Integrative Comp. Physiol. 296:R374-R382. (STA-320)

4. Rao, M. et al (2008). Mitochondrial DNA injury and mortality in hemodialysis patients. J. Am. Sco. Nephrol. 20:189-196. (STA-320)

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tion

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8-OHdG Levels in a Human Urine Sample.

OXIDATIVE & CELLULAR STRESS DNA / RNA Damage Assays


79

OxiSelect™ Comet Assays (Single Cell Gel Electrophoresis)


OxiSelect™ 3-Well Comet Assay Kit Light Microscopy

15 Wells STA-350

75 Wells STA-351


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


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.

• Versatile: High-level screening tool for DNA damage from a wide variety of sources

• User Friendly: Simple protocol with easy visu-alization by epifluorescence microscopy

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 was run under alkaline conditions at 33 V / 300 mA for 15 minutes.

Assay Principle for the OxiSelect™ Comet Assay Kit.



OxiSelect™ Oxidative DNA Damage Quantitation Kit (AP Sites)

80


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

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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.



81

Antioxidant Assays

OxiSelect™ Superoxide Dismutase Activity Assay

ROS generation is normally counterbalanced by the action of antioxidant enzymes and other redox molecules. We offer two types of assays for antioxidant quantitation:

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. 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. SOD activity is determined as the inhibition or reduction of chromagen.

• Sensitive: Detect as little as 0.6 units/mL • Fast: Obtain results in about 2 hours • Quantitative: SOD standard included as positive

control • Versatile: Suitable for use with urine, serum, cells

or tissue samples

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nm


OxiSelect™ Superoxide Dismutase Activity Assay Colorimetric 100 Assays STA-340

OxiSelect™ Catalase Activity Assay Kits


Colorimetric 96 Wells STA-341

Fluorometric 96 Wells STA-339 OxiSelect™ Catalase Activity Assay Kit

Catalase is a ubiquitous enzyme that destroys hydro-gen peroxides formed during oxidative stress. Our OxiSelect™ Catalase Activity Assay Kit provides a quick 30 minute protocol to monitor catalase activity from a variety of sample types. Kits are available with either colorimetric or fluorometric detection.

• Sensitive: Detect as little as 1.25 units/mL (colorimetric) or 50 mUnits/mL (fluorometric)

• Fast: Obtain results in less than 30 minutes • Versatile: Suitable for use with whole blood,

plasma, serum, cell lysates or tissue homoge-nates

• High Throughput: 96-well format

Standard Curve Using the OxiSelect™ Superoxide Dismu-tase Activity Assay.

OXIDATIVE & CELLULAR STRESS Antioxidant Assays

• Assays to quantify the presence of antioxidant enzymes • Assays to determine the antioxidant capacity of biomolecules


82

OxiSelect™ ORAC Activity Assay Kit


OxiSelect™ HORAC Activity Assay Kit Fluorometric 192 Assays STA-346

5 x 192 Assays STA-346-5

The assay known as ORAC (Oxygen Radical Anti-oxidant Capacity) is a powerful tool to measure the antioxidant capacity of biomolecules. The assay measures antioxidant capacity against peroxyl radi-cals. Our OxiSelect™ ORAC Activity Assay Kit measures antioxidant capacity quickly and easily in a variety of sample types.

• Sensitive: Detect as little as 2.5 µM • Fast: Obtain results in less than 2 hours on a

fluorometric plate reader • Versatile: Suitable for use with plasma, cell

fractions, tissue lysates, solid and aqueous nutrition samples

• Quantitative: Antioxidant standard included in each kit

Assay Principle for the OxiSelect™ Oxygen Radical Antioxidant Capacity (ORAC) Assay.

OXIDATIVE & CELLULAR STRESS Antioxidant Assays


OxiSelect™ ORAC Activity Assay Kit Fluorometric 192 Assays STA-345


OxiSelect™ HORAC Activity Assay Kit

The HORAC (Hydroxyl Radical Antioxidant Capac-ity) assay, like the ORAC assay above, is a powerful tool to measure the antioxidant capacity of bio-molecules. This assay specifically measures antioxi-dant capacity against hydroxyl radicals. Our OxiSelect™ HORAC Activity Assay Kit meas-ures antioxidant capacity quickly and easily in a vari-ety of sample types.

• Fast: Obtain results in less than 2 hours on a fluorometric plate reader

• Versatile: Suitable for use with plasma, cell fractions, tissue lysates, solid and aqueous nutrition samples

• Quantitative: Antioxidant standard included in each kit


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 in vitro and in vivo quantitation of various reactive oxygen species.

OXIDATIVE & CELLULAR STRESS ROS Assays

OxiSelect™ Cell-Based ROS Assay Kit

The OxiSelect™ ROS Assay Kit is a cell-based assay that measures the activity of hydroxyl, peroxyl, and other reactive oxygen species. The assay uses the cell-permeable fluorogenic 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. Fluo-rescence is quantified on a fluorometric plate reader.

• Sensitive: Detect concentrations as little as 10 pM

• Fast: Entire protocol takes about one hour • Versatile: Detects a wide variety of reactive

oxygen species

83


OxiSelect™ ROS Assay Kit Fluorometric 96 Assays STA-342

Assay Principle for the OxiSelect™ ROS Assay Kit.

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H2O2 Concentration

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Measurement of ROS in HeLa Cells. 50,000 HeLa cells in a 96-well plate were pretreated with 1mM DCFH-DA for 60 minutes at 37ºC. Cells were then treated with hydrogen peroxide for 20 minutes.


Recent Product Citation Zhang, Y. et al. (2010). The mitochondrial pathway of anesthetic isofluorane-induced apoptosis. J. Biol. Chem. 285:4025-4037. (STA-342)

OxiSelect™ Peroxide Detection Assays

OXIDATIVE & CELLULAR STRESS ROS Assays

84

Hydrogen peroxide is one of the most damaging re-active oxygen species. Our OxiSelect™ Peroxide Assay Kits provide a simple method for quantitation of hydrogen peroxide and other peroxides. Assays are available with either colorimetric or fluorometric detection. Our colorimetric assay measures the oxidation of ferrous (Fe2+) ions to ferric (Fe3+) ions in the pres-ence of peroxides. The ferric ions form a complex with a provided dye which may be read on a stan-dard microplate reader. The assay may be run with either aqueous phase or lipid phase samples. The fluorometric assay uses a probe which is con-verted from a non-fluorescent to a fluorescent state in the presence of peroxides and catalyzed by per-oxidases.

• Sensitive: Detect as little as 50 nM (fluorometric format) or 1 µM (colorimetric format)

• Fast: Easy 30 minute incubation • Versatile: Suitable for use with plasma, cell frac-

tions, tissue lysates, solid and aqueous nutrition samples

01000

2000300040005000

60007000

0 20 40 60

[H2O2] (µM)R

FU


Colorimetric 96 Wells STA-343

Fluorometric 96 Wells STA-344 OxiSelect™ Peroxidase Detection Assay


Standard Curve Generated with the OxiSelect™ Peroxide Detection Assay (Fluorometric).

86

CELL SIGNALING Cytoskeleton/Small GTPase

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:

Product Name Vectors Size Catalog Number

Cdc42 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 10 µg STA-455

GFP-Cdc42 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 10 µg STA-451

Rac1 Expression Vector Set Wild Type, T17N (Dom. Neg.), G12V (Active) 3 x 10 µg STA-454

GFP-Rac1 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 10 µg STA-450

Active Rac1 Expression Vector Set Q61L, Q61L/F37A, Q61L/Y40C 3 x 10 µg STA-458

H-Ras Expression Vector Set Wild Type, T17N (Dom. Neg.), G12V (Active) 3 x 10 µg STA-457

Active H-Ras Expression Vector Set V12, V12/S35, V12/C40 3 x 10 µg STA-459

RhoA Expression Vector Set Wild Type, T19N (Dom. Neg.), G14V (Active) 3 x 10 µg STA-456

GFP-RhoA Expression Vector Set Wild Type, T19N (Dom. Neg.), Q63L (Active) 3 x 10 µg STA-452


10 µg 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 re-searched small GTPases. Each set contains 3 vec-tors: wild type, dominant negative, and constitu-tively active forms. Vectors are supplied with or without a GFP reporter gene. Alternatively, choose active Rac1 or H-Ras vector sets (#STA-458 or STA-459). Each vector set con-tains 3 different active mutants.

Exoenzyme C3 (Rho Inhibitor) Expression Vector

Small GTPase Assays and Reagents

• Small GTPase Expression Vectors • Small GTPase Activation Assays

• Small GTPase Expression Vectors • Small GTPase Activation Assays


87


Small GTPase Activation Assays

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


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

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

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. Recent Product Citations

1. 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)

2. Harmon, B. et al (2008). Induction of the Gαq signaling cas-cade by the human immunodeficiency virus envelope is re-quired for virus entry. J. Virol. 82:9191-9205. (STA-400)

3. Lise, M-F. et al. (2009). Myosin-Va-interacting protein, RILPL2, controls cell shape and neuronal morphogenesis via Rac sig-naling. J. Cell Sci. 122:3810-3821. (STA-401-1)

4. Shen, E. et al. (2009). Rac1 is required for cardiomyocyte apoptosis during hyperglycemia. Diabetes 58:2386-2395. (STA-401-1)

5. Takano, A. et al. (2009). Identification of nectin-4 oncoprotein as a diagnostic and therapeutic target for lung cancer. Cancer Res. 69:6694-6703. (STA-401-1)

6. Zhang, S. et al (2008). The tumor suppressor LKB1 regulates lung cancer cell polarity by mediating Cdc42 recruitment and activity. Cancer Res. 68:740-748. (STA-402)

7. Pandey, D. et al. (2009). Unraveling a novel Rac1-mediated signaling pathway that regulates cofilin dephosphorylation and secretion in thrombin stimulated platelets. Blood 114:415-424. (STA-404)

8. Xu, Y. et al. (2010). Neuropilin-2 mediates VEGF-C-induced lymphatic sprouting together with VEGFR3. J. Cell Biol. 188:115-130. (STA-405)

9. Qu, Y. et al. (2008). Enhanced migration and CXCR4 over-expression in fibroblasts with telomerase reconstitution. Mol. and Cell. Biochem. 313:45-52. (STA-405)

10. Mohseni, M. et al (2008). The headpiece domain of dematin regulates cell shape, motility, and wound healing by modulating RhoA activity. Mol. Cell. Biol. 28:4712-4718. (STA-405)



88

Product Name Target Size Catalog Number

PAK1 PBD Agarose Beads Cdc42, Rac 400 µg STA-411

Raf1 RBD Agarose Beads Ras 400 µg STA-410

Rhotekin RBD Agarose Beads Rho 400 µg STA-412

Cdc42 G15A Agarose Beads Cdc42-GEF 800 µg STA-433

Rac1 G15A Agarose Beads Rac1-GEF 800 µg STA-432

RanBP1 Agarose Beads Ran 400 µg STA-421

RalBP1 PBD Agarose Beads Ral 400 µg STA-420

RhoA G17A Agarose Beads RhoA-GEF 400 µg STA-431

GGA3 PBD Agarose Beads Arf 400 µg STA-419

RalGDS RBD Agarose Beads Rap 400 µg STA-418

Small GTPase and GEF 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 and Active Rac-GEF Assay Kit.

Recent Product Citations 1. 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)

2. 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)

3. 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)

4. 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)

5. Sabbatini, M. et al (2008). Rap1 activation plays a regulatory role in pancreatic amylase secretion. J. Biol. Chem. 283:23884-23894. (STA-412)


Active Rac-GEF Assay Kit (Tiam1) Immunoblot/ECL 20 Assays STA-422

Visible Agarose Beads. Beads are easy to visualize, making it easier to avoid po-tential loss during washes and aspirations.

Active Rac-GEF Assay Kit (Tiam1)

Guanine nucleotide exchange factors (GEFs) activate small GTPases by catalyzing the exchange of GDP for GTP. Our Active Rac-GEF Assay Kit uses the agarose bead technology of our Small GTPase Activation Assays (see previous page) to pull down the active form of Rac-GEFs from endogenous lysates or purified samples. The specific GEF known as Tiam1 is then specifically detected with a polyclonal antibody.



89

Premade Recombinant Adenoviruses


Cdc42 ADV-152



Rac1 ADV-149





Ras V12C40 ADV-148

Ras V12S35 ADV-147



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

3. Rendon, B. et al. (2007). Regulation of human lung adenocar-cinoma cell migration and invasion by MIF. J. Biol. Chem. 282:29910-29918. (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)


6. Vaught, D. et al. (2009). Regulation of mammary gland branching morphogenesis by EphA2 receptor tyrosine kinase. Mol. Biol. Cell 20:2572-2581. (ADV-157)

7. Fang, W.B. et al. (2008). Overexpression of EPHA2 receptor destabilizes adherens junctions via a RhoA-dependent mechanism. J. Cell Sci. 121:358-368. (ADV-157)

8. Moldobaeva, A. et al. (2008). MIP-2 causes differential activa-tion of RhoA in mouse aortic versus pulmonary artery endo-thelial cells. Microvascular Res. 75:53-58. (ADV-157)

9. Brantley-Sieders, D. et al. (2008). The receptor tyrosine kinase EphA2 promotes mammary adenocarcinoma tumori-genesis and metastatic progression in mice by amplifying ErbB2 signal. J. Clin. Invest. 118:64-78. (ADV-157)

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.


Cdc42 L61 pBABEhygro RTV-203

K-Ras pBABEpuro RTV-220

K-Ras Q61 pWZLhygro RTV-221

myr-Rac1 pBABEpuro RTV-201

Rac1 V12 pBABEhygro RTV-202

N-Ras K61 pBABEpuro RTV-222

Rac3 V12 pBABEhygro RTV-205

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

Each vector is supplied as 100 µL of bacterial glycerol stock.

Gene-Specific Recombinant Retroviral Vectors

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


Generic Map of pBABEhygro Retroviral Expression Vector.

CELL SIGNALING Rho Kinase

90

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.

Our ROCK Activity Assays use a non-radioactive format to measure the level of active ROCK in cell or tissue lysates. The Immunoblot kit provides a convenient format for measuring ROCK activity in a few samples, while the ELISA kit contains a strip-well plate precoated with recombinant MYPT1 for higher throughput.


ROCK Activity Immunoblot Kit Immunoblot 20 Assays STA-415

96-Well ROCK Activity Assay Colorimetric 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-phosphor-MYPT1 as de-scribed in the assay protocol.

96-Well ROCK Activity Assay Principle.

Rho Kinase Assays

Recent Product Citations 1. Li, Z. et al. (2009). TrkBT1 induces liver metastasis of pancreatic

cancer cells by sequestering Rho GDP dissociation inhibitor and promoting RhoA activation. Cancer Res. 69:7851-7859. (STA-415)

2. Xiao, L. et al. (2009). ROCK mediates phorbol ester-induced apoptosis in prostate cancer cells via p21-Cip1 upr-egulation and JNK. J. Biol. Chem. 284:29365-29375. (STA-415)

3. Haas, B. et al. (2009). Protein kinase G controls brown fat cell differentiation and mitochondrial biogenesis. Sci. Signal. 2:ra78. (STA-416)


CELL SIGNALING GPCR Signaling

91

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 se-quence 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.

cAMP and cGMP 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 regula-tory molecules in the GPCR signaling cascade. Our cAMP and cGMP ELISA Kits provide a highly sen-sitive method to measure low levels of cAMP or cGMP in a variety of sample types. Sensitivity of these kits is very high, even with the colorimetric format.


cAMP ELISA Kit

Colorimetric 96 Assays STA-500


Chemiluminescent 96 Assays STA-501


cGMP ELISA Kit

Colorimetric 96 Assays STA-505


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, Colorimet-ric format.

0

0.5

1

1.5

2

2.5

0 20 40

Amount (ng)

OD

450

nm

PIP2

PIP3

PIP3 Binding to GRP-PH Domain.


92

CELL SIGNALING Reporter Assays & Stable Cell Lines

GFP Quantitation Kits


GFP ELISA Kit Colorimetric 96 Assays AKR-121

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 Quantitation Kit measures GFP fluo-rescence in a fluorometer, while our GFP ELISA kit uses a standard microplate reader.

Standard Curve Generated with the GFP ELISA Kit.

• Sensitive: Detection limit of 50 pg/ml with ELISA format

• Easy Quantitation: Measure GFP levels in a fluorometer or a standard microplate reader

ß-Galactosidase Staining Kit

LacZ is a commonly used reporter gene in transfection 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 efficient, easy-to-use method to determine the transfection effi-ciency of the LacZ gene. Each kit provides sufficient reagents to perform 75 assays in 35mm culture dishes.

0

0.5

1

1.5

2

2.5

3

3.5

0 200 400 600 800 1000

GFP (pg/mL)

OD

45

0 n

m


ß-Galactosidase Staining Kit 75 Assays AKR-100


293/GFP Cell Line 1 vial AKR-200

MDA-MB-231/GFP Cell Line 1 vial AKR-201

GFP Stable Cell Lines

These cell lines stably express green fluorescent protein.

Recent Product Citation 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. (AKR-100)

Recent Product Citations 1. 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. (AKR-120)

2. Rajan, S. et al. (2010). In vitro processing and secretion of mu-tant insulin proteins that cause permanent neonatal diabetes. Am. J. Physiol. Endocrinol. Metab. 298:E403-E410. (AKR-121)

3. Glenn, S.T. et al. (2008). In vivo analysis of key elements within the renin regulatory region. Physiol. Genomics 35:243-253. (AKR-121)


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 PhsophoBLOCKER™ Western Blot Blocking Reagent


Phospho Antibody Stripping Solution, 75X 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.

Rapid GST Inclusion Body Solubilization and Renaturation Kit


Rapid GST Inclusion Body Solubilization and Renaturation Kit 1 Kit AKR-110

Recombinant proteins expressed in bacteria often form inclusion bodies, especially when they are expressed at high levels. The Rapid GST Inclusion Body Solubilization and Renaturation Kit is de-signed to retrieve expressed GST fusion proteins in soluble form after lysis and extraction procedures. Each kit contains sufficient reagents to solubilize and renature up to 5-10 liters of bacterial culture.

• Faster Results: No lengthy dialysis or dilution steps

• Easy-to-Use: No pH variation or redox pair involved

• 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 dilu-tion; 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

Multiple Blotting and Stripping of 4G10 Phosphotyro-sine Antibody.

Recent Product Citation Ramakrishnan, R. et al. (2009). Characterization of Cdk9 T-loop phosphorylation in resting and activated CD4+ T lymphocytes. J. Leukoc. Biol. 86:1345-1350.

93

CELL SIGNALING Phosphorylation, Inclusion Bodies


Product Page 293 Cell Lines 35

AAV 38 Adenovirus 92 GFP Stable Expression 46 Lentivirus 55 Retrovirus 76

4-HNE Assay 76 8-Iso-Prostaglandin F2α ELISA 76 8-Isoprostane ELISA Kit 76 8-OHdG ELISA Kit (DNA Damage) 78 8-OHG ELISA Kit (RNA Damage) 78 AAV (Adeno-Associated Virus)

Cell Line 35 Expression & Packaging Systems 33-34 Expression Vectors 34 Purification Kit 35 Quantitation Kit 36 Titer Kit 36 Transduction Reagent 36

Active Rac-GEF Assay (Tiam1) 88 Adenovirus

Cell Line 38 Expression Systems 37, 69 Premade Recombinant 38-41 Purification Kits 42 Quantitation Kits 43 RCA Assay 44 Titer Kits 43 Transduction Reagent 44

Adhesion Assays 10-11 Advanced Glycation End

Products Assay 75 Advanced Oxidation Protein

Products Assay 75 AGE Assay 75 Alkaline Phosphatase Assays 30 Angiogenesis

Recombinant Adenovirus 38 Tube Formation Assay 22

Anoikis Assay 20 Antibody Stripping Solution 93 Antioxidant Assays

Catalase Activity Assay 81 HORAC Assay 82 ORAC Assay 82 Superoxide Dismutase Activity Assay 81

AOPP Assay 75 AP Sites Quantitation Kit 80 AUF1 Retroviral Vector 54 Autophagy Expression Vectors 22 β-Galactosidase

Recombinant Adenovirus 38 Recombinant Lentivirus 47 Reporter Assays 21, 92

Blocking Reagent 93 C3 Expression Vector 86 CA9 Recombinant Adenovirus 38 c-Abl Retroviral Vector 52 cAMP ELISA Kits 91 Cancer Cell Assays

Angiogenesis 22 Anoikis 20

Product Page

c-Myc Retroviral Vectors 53 Colony Formation Assays

Cell Transformation Assays 6-7 Hematopoietic Colony Forming

Cell Assay 28 Stem Cell Colony Formation

Assay 29 Tumor Cell Isolation Kit 9 Tumor Sensitivity Assay 8

Comet Assay Kits & Slides 79 Cre Recombinant Adenovirus 38 CSK Recombinant Adenovirus 41 Cyclic AMP ELISA Kits 91 Cyclic GMP ELISA Kits 91 CytoSelect™ Cell-Based Assays

Anoikis 20 Cell Adhesion 10-11 Cell Invasion 16-17 Cell Migration 12-18 Cell Transformation 6-7 Cell Viability 20 Chemotaxis 12-18 Colony Formation 6-9 Cytotoxicity 20 Haptotaxis 14 Phagocytosis 19 Soft Agar 6-9 Transmigration 15 Tumor Sensitivity 8 Wound Healing 18

Cytoskeleton Regulation Activation Assays 87 Adenoviruses 39 Expression Vectors 86 Retroviral Vectors 53

Cytotoxicity Assay 20 DCC Recombinant Adenovirus 39 DNA Damage Assays

8-OHdG ELISA Kit 78 AP Sites Quantitation Kit 80 Comet Assays 79 Double-Strand Break Assay 80

ECM (Extracellular Matrix) Kits Cell Adhesion Assays 10 Cell Invasion Assays 16-17 Tube Formation Assay 22

Endothelial Tube Assay 22 ERK2

Recombinant Adenovirus 40 Retroviral Vector 54

ERK5 Recombinant Adenovirus 40 ES/EC Cells

Alkaline Phosphatase Assays 30 Colony Formation Assays 29 Retroviral Expression Systems 26

Exoenzyme C3 Expression Vector 86 Feeder Cells 27 Firefly Luciferase Recombi-

nant Adenovirus 38 Fyn Recombinant Adenovirus 41 GEF (Guanine Exchange Factors)

Activation Assays 88 Agarose Beads 88

Clonogenic Tumor Cell Isolation 9

94

Product Page Cancer Cell Assays (cont’d)

Cell Adhesion 10-11 Cell Invasion 16-17 Cell Migration 12-18 Cell Transformation 6-7 Colony Formation 6-9 Soft Agar 6-9 Tumor Cell Isolation Kit 9 Tumor Sensitivity 8

Carbonyl Assays 74 Catalase Activity Assay 81 Cdc42

Activation Assay 87 Agarose Beads 88 Recombinant Adenovirus 39 Retroviral Vector 53

CEA Recombinant Adenovirus 38 Cell-Based Assays

Adhesion 10-11 Angiogenesis 22 Anoikis 20 Cell Contraction 21 Cell Viability 20 Chemotaxis 13, 17 Colony Formation 6-9 Cytotoxicity 20 Haptotaxis 14 Invasion 16-17 Migration 12-18 Phagocytosis 19 Senescence 21 Soft Agar 6-9 Transformation 6-7 Transmigration 15 Tumor Sensitivity 8 Wound Healing 18

Cell Cycle Adenoviruses 39 Anoikis Assay 20 Cell Viability Assay 20 Cytotoxicity Assay 20 Retroviral Vectors 52 Senescence Assays 21

Cell Invasion Assays 16-17 Cell Lines

293AAV 35 293AD 38 293LTV 46 293RTV 55 293/GFP 92 JK1 Feeder Cells 27 MDA-MB-231/GFP 92 MEF Feeder Cells 27 Plat-A Retroviral Packaging Cells 57 Plat-E Retroviral Packaging Cells 57 Plat-GP Retroviral Packaging Cells 57

SNL Feeder Cells 27 Cell Migration Assays 12-18 Cell Transformation Assays 6-7 Cell Viability Assay 20 cGMP ELISA Kits 91 Chemotaxis Assays 13, 17

PRODUCT INDEX—ALPHABETICAL


95

Product Page Lentivirus (cont’d)

Concentration & Purification Kits 50 Control Plasmids 47 Expression Systems 45-46 Expression Vectors 47 Packaging Systems 46 Premade Control Viruses 47 Purification Kits 50 Quantitation Kits 48-49 Titer Kits 48-49 Transduction Kits 51

Leukocyte Assays Adhesion 11 Transmigration 15

Lin-28 Retroviral Vectors 53 Lipid Peroxidation Assays

8-Isoprostane ELISA 76 HNE-His Adduct ELISA 76 MDA Assays 77 TBARS Assay 77

Luciferase Recombinant Adenovirus 38

MAP Kinase Signaling Recombinant Adenovirus 40 Retroviral Vectors 54

MAPKAPK2 Recombinant Adenovirus 40 Retroviral Vector 54

MDA Assay Kits 77 MDA-MB-231/GFP Stable

Cell Line 92 MEF Feeder Cells 27 MEK1


MEK5 Recombinant Adenovirus 40 MEKK1 Recombinant Adenovirus 40 MEKK3 Recombinant Adenovirus 40 microRNA Analysis

Adenoviral Expression System 69 Clone Collection 62-67 Control Vectors 68 Expression Vectors 62-68 Functional Reporter System 68 Lentiviral Expression System 69 Precursor Clone Collection 62-67 Retroviral Expression Vector 70 Transduction Enhancer 70

Migration Assays 12-18 miRNA Analysis

Adenoviral Expression System 69 Clone Collection 62-67 Control Vectors 68 Expression Vectors 62-68 Functional Reporter System 68 Lentiviral Expression System 69 Precursor Clone Collection 62-67 Retroviral Expression Vector 70 Transduction Enhancer 70

MKK3 Recombinant Adenovirus 40 Retroviral Vector 54

MKK4 Recombinant Adenovirus 40

PRODUCT INDEX—ALPHABETICAL Product Page GFP Products

ELISA Kit 92 Lentiviral Vectors 47 Quantitation Kits 92 Recombinant Adenovirus 38 Recombinant Lentivirus 47 Retroviral Vectors 52 Stable Cell Lines 92

GPCR Signaling Products 91 GST Inclusion Body Solubili-

zation and Renaturation Kit 93 GTPase Assay Kits 87 Haptotaxis Assays 14 HBV Core Antigen ELISA 60 HCV Core Antigen ELISA 60 HEK 293 Cell Lines

AAV 35 Adenovirus 38 GFP Stable Expression 92 Lentivirus 46 Retrovirus 55

Hematopoietic Colony Form-ing Cell Assay 28

Hepatitis B Core Antigen ELISA 60 Hepatitis C Core Antigen ELISA 60 HIF-1α Recombinant Adenovirus 40 HIV-1 p24 ELISA Kits 48-49 HNE-His Adduct ELISA Kit 76 hnRNAPA0 Retroviral Vector 54 HORAC Assay Kit 82 HuB Retroviral Vector 54 HuC Retroviral Vector 54 HuD Retroviral Vector 54 HuR Retroviral Vector 54 Hydrogen Peroxide Assays 84 Hydroxyl Radical Antioxidant

Capacity (HORAC) Assay 82 IFN Recombinant Adenovirus 40 IκB Recombinant Adenovirus 41 IKK Recombinant Adenovirus 41 IL-2 Recombinant Adenovirus 40 Immunoblot Blocking Reagent 93 In Vitro Angiogenesis Assay 22 In Vitro Tumor Sensitivity Assay 8 Inclusion Body Solubilization Kit 93 Induced Pluripotent Stem Cells

Lentiviral Vectors 25 Retroviral Packaging Cells 24 Retroviral Vectors 24

Invasion Assays 16-17 iPS Cell Reprogramming

Lentiviral Vectors 25 Retroviral Packaging Cells 24 Retroviral Vectors 24

JK1 Feeder Cells 27 JNK1


Klf4 Retroviral Vectors 53 KOSM Lentiviral Vector 25 LC3 Expression Vectors 22 Lentivirus

Cell Line 46

Product Page MKK6


MKK7 Recombinant Adenovirus 40 MyoD Recombinant Adenovirus 39 Myogenin Adenovirus 39 myr-Akt Retroviral Vectors 54 myr-Rac1


NANOG Retroviral Vectors 53 NFκB Adenoviruses 41 Nitrotyrosine Assay Kits 73 NOD2 Recombinant Adenovirus 41 NY-ESO-1 Adenovirus 38 Oct-3/4 Retroviral Vectors 53 ORAC Assay Kit 82 OxiSelect™ Oxidative

Stress Assays

Antioxidant Assays 81-82 DNA / RNA Damage Kits 78-80 Lipid Peroxidation Assays 76-77 Protein Oxidation Assays 73-75 ROS Assays 83-84

Oxygen Radical Antioxidant Capacity (ORAC) Assay 82

p24 ELISA Kits 48-49 p38

Recombinant Adenovirus 40 Retroviral Vectors 54

p53 Recombinant Adenovirus 39 Lentiviral Vectors (shRNA) 25 Retroviral Vectors 52

p68 RNA Helicase Adenovirus 39 PABP Retroviral Vector 54 PAK1

PBD Agarose Beads 88 Recombinant Adenovirus 39

Peroxide Detection Assays 84 Phagocytosis Assays 19 Phospho Antibody Stripping

Solution 93 PhosphoBLOCKER™ Western

Blot Blocking Reagent 93 PI3K Retroviral Vector 54 PKC Recombinant Adenovirus 41 Plat-A Retroviral Packaging

Cells 57 Plat-E Retroviral Packaging Cells 57 Plat-GP Retroviral Packaging

Cells 57 Platinum Retroviral Expression

Expression Systems 56 Packaging Cell Lines 57

PRAK Recombinant Adenovirus 40 Retroviral Vector 56

Protease Retroviral Vectors 53 Protein Oxidation Assays

Advanced Glycation End Prod-ucts (AGE) 75


Product Page Protein Oxidation Assays (cont’d)

Advanced Oxidation Protein Products (AOPP) 75

Carbonyl 74 Nitrotyrosine 73

QuickTiter™ Viral Quantitation & Titer Kits AAV 36 Adenovirus 43 Lentivirus 48-49 Retrovirus 59

Rac Activation Assay 87 Agarose Beads 88 GEF Assay 88 Recombinant Adenovirus 39 Retroviral Vectors 53

Raf1 Recombinant Adenovirus 40 Retroviral Vectors 54

Ral Activation Assay 87 Agarose Beads 88

Ran Activation Assay 87 Agarose Beads 88

RAPAd® Adenoviral Expression Systems 37, 69

Rapid GST Inclusion Body Solu-bilization and Renaturation Kit 93

Rapid RCA Assay 44 Ras Superfamily

Activation Assays 87 Agarose Beads 88 Expression Vectors 86 Recombinant Adenovirus 39 Retroviral Vectors 53

RCA Assay Kit 44 Reactive Oxygen Species (ROS)

Assays 83-84 Recombinant Adenoviruses 38-41 Recombinant GRP-PH Domain 91 Rel B Recombinant Adenovirus 41 Reporter Genes

Lentiviral Vectors 47 Quantitation Assays 92 Recombinant Adenovirus 38 Recombinant Lentivirus 47 Retroviral Vectors 52 Stable Cell Lines 92

Retrovirus Concentration & Purification Kits 58 Expression Systems 56 Expression Vectors 55 Gene-Specific Vectors 52-54 Packaging Cell Lines 55, 57 Purification Kits 58 Quantitation Kits 59 Transduction Kits 60

Rho Activation Assay 87 Agarose Beads 88 Recombinant Adenovirus 39 Retroviral Vector 53

Product Page Rho Kinase Activity Assays 90 RNA Damage ELISA Kit 78 RNAi Enhancer Reagent 70 ROCK Activity Assay Kits 90 ROS Assays 83-84 SCGE Assay Kits 79 SEAP Recombinant Adenovirus 38 Senescence Assays 21 shAkt1 Recombinant Adenovirus 41 shAkt2 Recombinant Adenovirus 41 Single Cell Gel Electrophoresis

Assays 79 Small GTPase

Activation Assays 87 Active GEF Assays 88 Agarose Beads 88 Expression Vectors 86 Premade Adenoviruses 39 Retroviral Vectors 53

SNL Feeder Cells 27 SOD Activity Assay Kit 81 Soft Agar Colony Formation Kits

Cell Transformation Assays 6-7 Hematopoietic Colony Forming

Cell Assay 28 Stem Cell Colony Formation

Assay 29 Tumor Cell Isolation Kit 9 Tumor Sensitivity Assay 8

SOK Recombinant Adenovirus 40 Sox2 Retroviral Vector 53 Src Recombinant Adenovirus 41 Stat5 Retroviral Vectors 54 Stem Cell Research

Alkaline Phosphatase Detection Kits 30

Feeder Cells 27 Hematopoietic Colony Forming

Cell Assay 28 iPS Cell Reprogramming 24-25 PCR Primers 30 Retroviral Expression Systems 26 Stem Cell Colony Formation

Assay 29 Total Protein—Murine ES Cell

Line D3 30 Total RNA—Murine ES Cell Line

D3 30 Superoxide Dismutase Assay 81 Tac-Rac1 Adenovirus 40 TBARS Assay Kit 77 TIA1 Retroviral Vector 54 Tiam1 Assay Kit 88 TIAR Retroviral Vector 54 Total Protein—Ms ES Cell Line D3 30 Total RNA—Ms ES Cell Line D3 30 Transcription Regulation

Retroviral Vectors 54 Transformation Assays 6-7 Transmigration Assays 15 TTP Retroviral Vector 54 Tube Formation Assay 22 Tumor Antigen Adenoviruses 38

Product Page Tumor Cell Assays

Cell Adhesion 10-11 Cell Invasion 16-17 Cell Migration 12-18 Cell Transformation 6-7 Chemosensitivity 8 Soft Agar Colony Formation 6-9 Transmigration 15 Tumor Cell Isolation 9

Tyrosine Kinase Adenoviruses 41 uPA / uPAR Retroviral Vectors 53 VEGF Recombinant Adenovirus 38 ViraBind™ Purification Kits

AAV 35 Adenovirus 42 Lentivirus 50 Retrovirus 58

ViraDuctin™ Transduction Kits & Reagents

AAV 36 Adenovirus 44 Lentivirus 51 Retrovirus 60

Viral Expression Systems AAV 33-34 Adenovirus 37 Lentivirus 45-46 Retrovirus 56

Viral Packaging Cells AAV 35 Adenovirus 38 Lentivirus 46 Retrovirus 55

Viral Titer Kits AAV 36 Adenovirus 43 Lentivirus 48-49 Retrovirus 59

Viral Transduction Reagents AAV 36 Adenovirus 44 Lentivirus 51 Retrovirus 60

ViraSafe™ Lentivirus Expression Systems

45-46, 69

Virus Purification Kits AAV 35 Adenovirus 42 Lentivirus 50 Retrovirus 58

Virus Quantitation Kits AAV 36 Adenovirus 43 Lentivirus 48-49 Retrovirus 59

VSV-G Retroviral Vector 55 Western Blot Blocking Reagent 93 Wound Healing Assay 18

PRODUCT INDEX—ALPHABETICAL


97

PRODUCT INDEX—CATALOG NO.

Catalog No. Page AAV-100 35 AAV-200 36 AAV-201 36 AAV-400 through AAV-402

34

AD-100 38 AD-200 44 AD-201 44 ADV-001 through ADV-008

38

ADV-100 40 ADV-101 38 ADV-102 through ADV-164

40

ADV-202 through ADV-210

39


41


39


38

AKR-100 92 AKR-102 through AKR-110

93

AKR-120 through AKR-201

92

CBA-050 through CBA-071

10

CBA-080 20 CBA-081 20 CBA-100 13 CBA-100-C 17 CBA-100-COL 14 CBA-100-FN 14 CBA-101 13 CBA-101-C 17 CBA-101-COL 14 CBA-101-FN 14 CBA-102 through CBA-106

13

CBA-106-C 17 CBA-110 through CBA-112-LN

17

CBA-120 18

Catalog No. Page CBA-120-5 18 CBA-130 6 CBA-130-5 6 CBA-135 through CBA-145-5

7

CBA-150 8 CBA-150-5 8 CBA-155 9 CBA-155-5 9 CBA-200 22 CBA-201 21 CBA-210 11 CBA-211 11 CBA-212 15 CBA-215 11 CBA-216 15 CBA-220 19 CBA-224 19 CBA-230 21 CBA-231 21 CBA-240 20 CBA-300 through CBA-305

30

CBA-310 through CBA-316

27

CBA-320 28 CBA-320-5 28 CBA-325 29 CBA-325-5 29 LTV-100 46 LTV-200 51 LTV-201 51 LTV-300 through LTV-403

47

LTV-451 25 LTV-700 25 LTV-801 22 MIR-1-1 through MIR-30C-2

62

MIR-30D through MIR-152

63

MIR-154 through MIR-302B

64

MIR-302C through MIR-491

65

MIR-499 through MIR-671

66

Catalog No. Page

MIR-676 through

67

MIR-EXP-C 68 MIR-EXP-GP-C 68 MIR-GFP 68 MIR-LET7A-2 through MIR-LET7I

62

MIR-NULL 68 MIR-NULL-GP 68 MMU-LET7A-1 through MMU-LET7G

62

MMU-MIR-1-1 through MMU-MIR-30C-1

62

MMU-MIR-30C-2 through MMU-MIR-151

63

MMU-MIR-152 through MMU-MIR-302A

64

MMU-MIR-302B through MMU-MIR-497

65

MMU-MIR-499 through MMU-MIR-675

66

MMU-MIR-676 through MMU-MIR-1907

67

RNA-200 70 RNA-201 70 RTV-001-HYGRO

55

RTV-001-NEO 55 RTV-001-PURO

55

RTV-002 52 RTV-003 through RTV-041

55

RTV-050 52 RTV-051 52 RTV-052 52 RTV-101 through RTV-104

53

RTV-105 through RTV-125

54


53


54

Catalog No. Page RTV-400 24 RTV-401 through RTV-405

52

RTV-410 24 RTV-501 through RTV-712

53

RTV-801 22 RV-100 55 RV-101 57 RV-102 57 RV-103 57 RV-110 through RTV-114

55

RV-200 60 RV-201 60 STA-200 91

STA-200-1MG

91

STA-303 through STA-305-5

73

STA-308 through STA-315

74

STA-316 through

75

STA-320 78 STA-320-5 78 STA-321 80 STA-324 80 STA-325 78 STA-325-5 78 STA-330 through STA-333

77

STA-334 76 STA-335 76 STA-337 76 STA-339 81 STA-340 81 STA-341 81 STA-342 83 STA-343 83 STA-344 83 STA-345 82 STA-346 82 STA-350 through STA-356-5

79


87

Catalog No. Page


86

STA-500 through STA-506-5

91

VPK-090 through VPK-096-5

50

VPK-099 42 VPK-100 42 VPK-101 42 VPK-106 43 VPK-107 through VPK-108-HIV-P24-5

49

VPK-109 43 VPK-110 43 VPK-111 44 VPK-112 49 VPK-120 59 VPK-130 through VPK-136-5

58

VPK-140 through VPK-141-5

35

VPK-145 36 VPK-150 60 VPK-151 60 VPK-205 46 VPK-206 46 VPK-211 through VPK-220-PAN

46-47

VPK-250 through VPK-253

37

VPK-300 through VPK-308

57

VPK-402 through VPK-419-SER2

34


88

STA-410 88 STA-411 88 STA-412 88 STA-415 90 STA-416 90


WORLDWIDE DISTRIBUTORS

UNITED STATES Cell Biolabs, Inc. Tel: +1 858 271-6500 Fax: +1 858 271-6514 [email protected] www.cellbiolabs.com

CANADA Cedarlane Laboratories Ltd. Tel: 905-878-8891 Toll-Free: 800-268-5058 Fax: 905-878-7800 [email protected] www.cedarlanelabs.com

North America

Africa

EGYPT New Test Co. (NTCO) Tel: +20 3 358 3543 Fax: +20 3 359 6836 [email protected]

SOUTH AFRICA Biocom Biotech Tel: +27 12 654 4614 Fax: +27 86 654 9048 [email protected] www.biocombiotech.com.za

Asia

AUSTRALIA Jomar Bioscience Pty. Ltd. Tel: +61 8 8362 6766 Fax: +61 8 8362 6388 [email protected] www.jomar.on.net

BAHRAIN DPC Lebanon S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected] www.dpcleb.com

CHINA (P.R.C.) Beijing Seajet Scientific Co. Tel: +86 10 8859-7838 Fax: +86 10 8859-5011 [email protected] www.seajetsci.com

Genetimes Technology, Inc. Tel: +86 21 5426-2677 Fax: +86 21 6439-8855 [email protected] www.genetimes.com.cn

HONG KONG Beijing Seajet Scientific Co. [email protected] www.seajetsci.com

Genetimes Technology Tel: +852 2385 2818 Fax: +852 2385 1308 [email protected] www.genetimes.com.hk

INDIA Biogenuix Medsystems Pvt. Ltd. Tel: +91-9810168857 Fax: +91-11-41658590 [email protected] www.biogenuix.com

GenxBio Health Sciences Pvt. Ltd. Tel: +91-11-64608275 Fax: +91-11-43021545 [email protected] www.genxbio.com

INDONESIA Precision Technologies Pte Tel: +65 6273 4573 Fax: +65 6273 8898 [email protected] www.pretech.com.sg

IRAQ DPC Lebanon S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected] www.dpcleb.com

ISRAEL BioConsult Tel: +972-2-5667043 Fax: +972-2-5662790 [email protected] www.bioconsult.co.il

JAPAN Cosmo Bio Co., Ltd. Tel: +81 3 5632 9610 Fax: +81 3 5632 9614 [email protected] www.cosmobio.co.jp

JORDAN DPC Lebanon S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected] www.dpcleb.com

KOREA CMI Biotech Tel: 82-2-444-7101 Fax: 82-2-444-7201 [email protected] www.cmibio.com

Koma Biotech Inc. Tel: 82-2-579-8787 Fax: 82-2-578-7042 [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] www.dpcleb.com

LEBANON DPC Lebanon S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected] www.dpcleb.com

MALAYSIA Precision Technologies Pte Ltd. Tel: +65 6273 4573 Fax: +65 6273 8898 [email protected] www.pretech.com.sg

NEW ZEALAND

Jomar Bioscience Pty. Ltd. Tel: +61 8 8362 6766 Fax: +61 8 8362 6388 [email protected] www.jomar.on.net

OMAN DPC Lebanon S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected] www.dpcleb.com

PAKISTAN Madina Scientific Stores Tel: +92 42 7248361 Fax: +92 42 7314122 [email protected]

QATAR DPC Lebanon S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected] www.dpcleb.com

SAUDI ARABIA DPC Lebanon S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected] www.dpcleb.com

SINGAPORE Precision Technologies Pte Ltd. Tel: +65 6273 4573 Fax: +65 6273 8898 [email protected] www.pretech.com.sg

TAIWAN GeneTeks BioScience, Inc. Tel: 886-2-2970 1170 Fax: 886-2-970 1172 [email protected]

HoHo Technology Co., Ltd. Tel: 02-2748-9697 Fax: 09-4329-5989 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

UNITED ARAB EMIRATES DPC Lebanon S.A.R.L. Tel: 00961 1 502 812 Fax: 00961 8 805 851 [email protected] www.dpcleb.com

BRAZIL Sellex (S.A.C.) Tel: +55 (011) 5506-4646 Fax: +55 (011) 5505-7433 [email protected] www.sellex.com

CHILE Fermelo S.A. Tel: +56 2 247 2976 Fax: +56 2 247 2977 [email protected] www.fermelo.com

South America


WORLDWIDE DISTRIBUTORS

ALBANIA Pharmex SA Tel: 0030 210 5199200 Fax: 0030 210 5144279 [email protected] www.pharmex.gr

AUSTRIA THP Medical Products Vertriebs GmbH Tel: +43 (1) 292 82 80 Fax: +43 (1) 292 82 80 88 [email protected] www.thp.at

BioCat GmbH Tel: +49 (0) 6221 7141516 Fax: +49 (0) 6221 7141529 [email protected] www.biocat.com

BELGIUM Bio-Connect BV Tel: +31 (0)26 326 4450 Fax: +31 (0)26 326 4451 [email protected] www.bio-connect.nl

CROATIA Interos2000 Ltd. Tel: +385 1 617 0059 Fax: +385 1 617 0086 [email protected] www.rotaja.com

CZECH REPUBLIC THP Medical Products Vertriebs GmbH Tel: +43 (1) 292 82 80 Fax: +43 (1) 292 82 80 88 [email protected] www.thp.at

DENMARK Bio-Mediator Ky Tel: +358 9 852 4898 Fax: +358 9 852 4884 [email protected] www.bio-mediator.com

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-mediator.com

FRANCE Euromedex Tel: +33 3 88 18 07 27 Fax: +33 3 88 18 07 28 [email protected] www.euromedex.com

GERMANY


THP Medical Products Vertriebs GmbH Tel: +43 (1) 292 82 80 Fax: +43 (1) 292 82 80 88 [email protected] www.thp.at

GREECE Pharmex SA Tel: 0030 210 5199200 Fax: 0030 210 5144279 [email protected] www.pharmex.gr

HUNGARY THP Medical Products Vertriebs GmbH Tel: +43 (1) 292 82 80 Fax: +43 (1) 292 82 80 88 [email protected] www.thp.at

IRELAND Cambridge Bioscience Ltd. Tel: +44 (0)1223 316 855 Fax: +44 (0)1223 360 732 [email protected] www.bioscience.co.uk

ITALY Valter Occhiena S.R.L. Tel: +39 011 771 69 71 Fax: +39 011 776 18 00 [email protected] www.valterocchiena.com

LATVIA Upstream OU Tel: +372 51 86 295 [email protected] www.upstream.ee

LITHUANIA 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] www.bio-connect.nl

MACEDONIA Pharmex SA Tel: 0030 210 5199200 Fax: 0030 210 5144279 [email protected] www.pharmex.gr

NETHERLANDS

Bio-Connect BV Tel: +31 (0)26 326 4450 Fax: +31 (0)26 326 4451 [email protected] www.bio-connect.nl

NORWAY Bio-Mediator Ky Tel: +358 9 852 4898 Fax: +358 9 852 4884 [email protected] www.bio-mediator.com

POLAND STI Tel: +48 (61) 6417759 Fax: +48 (61) 6417758 [email protected] www.sti.biz.pl

SERBIA Swiss Concept Ltd. Tel: +381 11 3610 945 Fax: +381 11 3610 945 [email protected]

SLOVAKIA THP Medical Products Vertriebs GmbH Tel: +43 (1) 292 82 80 Fax: +43 (1) 292 82 80 88 [email protected] www.thp.at

SLOVENIA THP Medical Products Vertriebs GmbH Tel: +43 (1) 292 82 80 Fax: +43 (1) 292 82 80 88 [email protected] www.thp.at

SPAIN bioNova Cientifica S.L. Tel: +34 91 551 54 03 Fax: +34 91 433 45 45 [email protected] www.bionova.es

Europe

SWEDEN Bio-Mediator Ky Tel: +358 9 852 4898 Fax: +358 9 852 4884 [email protected] www.bio-mediator.com

SWITZERLAND LuBioScience GmbH Tel: +41 41 417 02 80 Fax: +41 41 417 02 89 [email protected] www.lubio.ch


TURKEY

MedSanTek Ltd. Co. Tel: +90 212 635 85 46 Fax: +90 212 635 83 50 [email protected] www.medsantek.com.tr

UNITED KINGDOM

Cambridge Bioscience Ltd. Tel: +44 (0)1223 316 855 Fax: +44 (0)1223 360 732 [email protected] www.bioscience.co.uk

www.cellbiolabs.com [email protected] 99

2010 cell biolabs catalog final

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