cell biolabs catalog web

8/10/2019 Cell Biolabs Catalog WEB

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Cell-Based Assays

Ordering Information & Support 4

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Stem Cell Research 25

Viral Expression 33

microRNA Analysis 65

TRANSFORMATION ADHESION MIGRATION INVASION WOUND H EALING

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

TABLE OF C ONTENTS

PHAGOCYTOSIS VIABILITY/D EATH SENESCENCE CONTRACTION ANGIOGENESIS

IPS C ELL REPROGRAMMING RETROVIRAL EXPRESSION SYSTEMS COLONY FORMATION A SSAYS ALKALINE P HOSPHATASE A SSAYS FEEDER C ELLS

ADENOVIRUS ADENO -ASSOCIATED

VIRUS (AAV)LENTIVIRUS RETROVIRUS

PREMADE VIRUSES EXPRESSION SYSTEMS PURIFICATION KITS T ITER KITS TRANSDUCTION KITS

PRECURSOR C LONE C OLLECTION MAMMALIAN EXPRESSION VECTORS VIRAL EXPRESSION P LASMIDS FUNCTIONAL REPORTER SYSTEM KNOCKDOWN ENHANCER


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Oxidative Stress / Damage 75

Cell Signaling & Protein Biology 97

Product IndexWorldwide Contacts

123130

TABLE OF C ONTENTS

www.cellbiolabs.com [email protected]

Metabolism Research 111

Infectious Disease Research 119

PROTEIN O XIDATION /N ITRATION LIPID P EROXIDATION DNA/RNA D AMAGE AND REPAIR REACTIVE O XYGEN SPECIES (ROS) A SSAYS ANTIOXIDANT A SSAYS

SMALL GTP ASE / G-P ROTEIN S IGNALING KINASE A SSAYS REPORTER C ELLS AND REAGENTS EPITOPE T AGS PROTEIN P HOSPHORYLATION

CHOLESTEROL A SSAYS APOLIPOPROTEIN A SSAYS APOLIPOPROTEINS AND LIPOPROTEINS ANTIBODIES TO A POLIPOPROTEINS RENAL FUNCTION A SSAYS

VIRUS C ORE A NTIGEN A SSAYS BACTERIAL RAPID D ETECTION KITS


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ORDERING INFORMATION & S UPPORT

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Worldwide Technical SupportPlacing an Order within the U.S.

Placing an Order outside the U.S.

Our Customer Service representatives areavailable Monday through Friday from 8 amto 5 pm Pacific Time.

Most orders received by 2 pm Pacific Timewill be shipped the same day. We will notifyyou immediately of any backordered item.

We accept VISA, MasterCard and Ameri-can Express cards. Net 30 day terms maybe offered upon credit approval.

Phone 1 858 271 65001 888 CBL 0505 (Toll-Free)

Fax 1 858 271 6514

E-mail [email protected]

Online www.cellbiolabs.com

Mail Attn: Customer Service7758 Arjons DriveSan Diego, CA 92126

Pricing

Kit Components

Do you have questions about a particularproduct before you buy? Do you need helpwith a protocol?

Our Technical Service Scientists have beendirectly involved in the development andtesting of our products, so you get the bene-fit of their hands-on experience.

Phone 1 858 271 65001 888 CBL 0505 (US Toll-Free)

Fax 1 858 271 6514

E-mail [email protected]

Current U.S. prices are available online atwww.cellbiolabs.com , or you may request aseparate price list by e-mail. Prices are sub-

ject to change without notice.

For pricing outside the U.S. please contactyour local distributor, which can be found onthe inside back cover of this catalog.

Because our kits are QC tested by lot, indi-vidual kit components are generally notavailable for purchase separately. However,certain components may be available in

bulk quantities on a custom basis. For moreinformation please inquire by sending amessage to [email protected] .

Please see our Worldwide Distributorssection on the inside back cover of this cata-log. We have a network of global distributorsserving life science researchers in over 70

countries.

If you dont see your country listed, pleasecontact our U.S. office.

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


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CytoSelect 96-Well Cell Transformation AssayTraditionalSoft Agar Colony Formation

Product Name Detection Size Catalog Number

CytoSelect 96-Well Cell Transformation Assay (Soft Agar ColonyFormation) Fluorometric

1 Plate* CBA-130

5 Plates* CBA-130-5

Our CytoSelect 96-Well Cell Transformation Assay(Soft Agar Colony Formation) is suitable for measur-ing malignant transformation where no downstreamanalysis is required. Transformed cells cannot be re-covered; however, no manual cell counting is re-quired.

With this assay, cells are incubated in a semisolidagar medium for 6-8 days, then solubilized, lysed anddetected using CyQuant GR dye in a fluorometricplate reader.

Fast Results : 6-8 days vs. 21 daysPlate Reader Convenience : Eliminates manualcountingVersatile Format : Designed for 96-well through-put, but can be adapted for 48, 24, 12 or 6-well

CELL-BASED A SSAYS 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, allowhigh-throughput drug screening, and enable recovery of transformed cells for downstreamanalysis. These advances have also allowed us to develop a unique kit for the separationof clonogenic cancer cells from normal cells in heterogeneous solid tumors.

Tumor Cell / Soft Agar Assays

Cell Transformatio n Assay Principl e.

Recent Product Citations

1. Inami, Y. et al. (2011). Persistent activation of Nrf2 through p62in hepatocellular carcinoma cells. J. Cell Biol. 193(2): 275-284.

2. Carnahan, J. et al. (2010). Selective and potent Raf inhibitorsparadoxically stimulate normal cell proliferation and tumorgrowth. Mol. Cancer Ther . 9:2399-2410.

3. Liu, F. et al. (2010). Epigenomic alterations and gene expres-sion profiles in respiratory epithelia exposed to cigarette smokecondensate. Oncogene 29 :3650-3664.

4. Iorns, E. et al. (2010). The role of SATB1 in breast cancerpathogenesis. J. Natl. Cancer Inst. 102(16): 1284-1296.

5. Faoro, L. et al. (2010). EphA2 mutation in lung squamous cellcarcinoma promotes increased cell survival, cell invasion, focaladhesions, and mammalian target of rapamycin activation. J.Biol. Chem. 285 :18575-18585.

6. Rubio, R. et al. (2010). Deficiency in p53 but not retinoblastomainduced the transformation of mesenchymal stem cells in vitroand initiates leiomyosarcoma in vivo. Cancer Res . 70 :4185-4194.

7. 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 immunocompetentmice. Mol. Cancer Ther. 8:1692-1701.



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CytoSelect 96-Well Cell Transformation AssaysAdvancedSoft Agar with Post-Incubation Cell RecoveryThe CytoSelect 96-Well Cell Transformation Assay(Cell Recovery Compatible) provides a robust systemfor 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 daysCell Recovery : Transformed cells remain viablefor further analysisPlate Reader Convenience : Eliminates manualcounting of cellsVersatile Format : Designed for 96-well through-put, but can be adapted for 48, 24, 12 or 6-well

Cell Transformatio n Assay Principl e. Cell colonies form after a 6-8 day incubation with agar matrix. Transformed cells can then beeither lysed and detected with a fluorescent dye or recovered andre-plated.

Easy Fluorescence Detection with the CytoSelect Cell Trans-formation Assay. HeLa and NIH3T3 cells were seeded at variousconcentrations and cultured for 6 days. Transformed colonies werequantified 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)

Colorimetric1 Plate* CBA-135

5 Plates* CBA-135-5

Fluorometric1 Plate* CBA-140

5 Plates* CBA-140-5

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

5 Plates*** CBA-145-5Fluorometric


Recent Product Citations 1. Mathew, B. et al. (2011). The novel role of the mu opioid recep-

tor in lung cancer progression: A laboratory investigation. Anesth. Analg. 112 :558-567. (CBA-135)

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

3. Hirata, H. et al. (2009). Wnt antagonist gene DKK2 is epigeneti-cally silenced and inhibits renal cancer progression throughapoptotic and cell cycle pathways. Clin. Cancer Res. 15 :5678-5687. (CBA-135)

4. Xie, G. et al. (2009). Acetylcholine-induced activation of M3muscarinic receptors stimulates robust matrix metalloproteinasegene expression in human colon cancer cells. Am. J. Physiol.Gastrointest. Liver Physiol. 296: G755-G763. (CBA-135)



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CytoSelect 96-Well In Vitro Tumor Sensitivity Assay Colorimetric96 Assays CBA-150

5 x 96 Assays CBA-150-5

CytoSelect 96-Well In Vitro Tumor Sensitivity Assay

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Inhibitio n of HeLa Cell Transformation by 5-Fluorouracil . HeLacells were seeded at 5000 cells/well and cultured 7 days at various5-FU concentrations. Cell transformation was determined accord-ing to the assay protocol. IC50 value of 5-Fluorouracil on HeLa cellanchorage-independent growth was determined to be ~ 1 M.

Inhibitio n of HeLa Cell Anc horage-IndependentGrowth by Taxol . HeLa cells were cultured for 7days in the absence (top) or presence (bottom) of 1nM Taxol according to the assay protocol.

The CytoSelect In Vitro Tumor Sensitivity Assayprovides a stringent, anchorage-independent modelfor chemosensitivity testing and possible anticancerdrug screening. The assay uses a soft agar matrix topromote the colony formation of neoplastic cells inabout a week. Cells are quantified using a standardELISA plate reader.

Fast Results : 6-8 daysIn Vivo Simulation : Resembles a three-dimensional cell environmentPlate Reader Convenience : Eliminates manualcounting

Tumor Sensiti vity Assay Principle.

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Recent Product Citations 1. Itamochi, H. et al. (2011). Inhibiting the mTOR pathway syner-

gistically enhances cytotoxicity in ovarian cancer cells inducedby etoposide through upregulation of c-Jun. Clin. Cancer Res.17 :4742-4750.

2. Kang, D.W. et al. (2010). Phospholipase D1 drives a positivefeedback loop to reinforce the Wnt/-catenin/TCF signalingaxis. Cancer Res. 70 :4233-4242.



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CytoSelect Clonogenic Tumor Cell Isolation Kit

Product Name Size Catalog Number

CytoSelect Clonogenic Tumor Cell Isolation Kit5 Preps CBA-155

25 Preps CBA-155-5

Clonogenic Colon y Formation, Isolation and Re-plating. A:Clonogenic colony formation (red arrows) and single cells (blackarrows) after 7 day incubation. B: Isolation of clonogenic coloniesfrom single cells. C: Re-plated clonogenic colonies after 3 days (notrypsinization). D: Re-plated clonogenic colonies 1 day aftertrypsinization.

Clean separation of clonogenic tumor cells from nor-mal cells is critical for proper analysis of disease stateprogression. Due to the heterogeneity of many tu-mors, however, isolation of homogenous tumor cellpopulations can be difficult.

The CytoSelect Clonogenic Tumor Cell Isolation Kituses a proprietary semisolid agar medium to facilitatecolony formation by cells from solid tumors.

Colonies are grown in either a 6-well plate or a 35mm dish. The colonies are then isolated away fromsingle cells by size filtration.

Efficient : Easily eliminates single cells fromclonogenic tumor cell population Versatile: In addition to solid tumors, haspotential use in isolating tumor stem cells

Clonogenic Tumor Cell Isolation Procedure.




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CytoSelect ECM Cell Adhesion Assays


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

Colorimetric 48 Assays CBA-070

Fluorometric 48 Assays CBA-071

CytoSelect 48-Well Cell Adhesion Assay, Collagen IColorimetric 48 Assays CBA-052


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


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


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


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


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

The CytoSelect ECM Cell Adhesion Assays providea quantitative method for evaluation of cell adhesion.The 48-well plate is precoated with your choice ofsubstrate. Cells are seeded onto the substrate; adher-ent cells attach, while non-adherent cells are washed

away. Adherent cells can be quantified on a standardplate reader or fluorometer.

Quantitative : Measure results in a colorimetric orfluorescence plate readerFlexible : Uniform substrate layer of your choice ofCollagen I, Collagen IV, Fibrinogen, Fibronectin, orLaminin; or choose the ECM array which containsall 5 ECM proteins

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CELL-BASED A SSAYS Cell Adhesion

BSA

Vitronectin

Laminin

Fibronectin

Collagen IV

Collagen I

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

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

MDA-231 HT-1080 HEK293Recent 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 fromnormal African Americans and Caucasian Americans do-nors. PLoS One 3(8) :E2847. (CBA-060)



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CytoSelect Leukocyte Endothelium Adhesion Assays

Product Name Detection Size Catalog NumberFluorometric 96 Assays CBA-210


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

CytoSelect Leukocyte-Endothelium Adhesion Assay

CytoSelect Leukocyte-Epithelium Adhesion Assay

CytoSelect Leukocyte-endothelium Adhesio n AssayPrinciple.

Human Monocytic THP-1 Adhesion to HUVEC MonolayerUsing the CytoSelect Leukocyte-endothelium Adh esion

Ass ay. HUVEC monolayer was treated with 1 M PMA for 12 hrs.LeukoTracker labeled THP-1 cells were allowed to attach toHUVEC monolayer for 1 hr. Adherent cells were lysed and quan-tified as described in the assay protocol.

The CytoSelect Leukocyte Endothelium Adhesion Assays provide a robust system for the quantitativedetermination of interactions between leukocytesand endothelium. Adherent cells can be quantifiedon a fluorescence plate reader.

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CELL-BASED A SSAYS Cell Adhesion

Recent Product Citations 1. Xue, J. et al. (2009). NFkB regulates thrombin-induced ICAM-

1 gene expression in cooperation with NFAT by binding to theintronic NFkB site in the ICAM-1 gene. Physiol. Genomics38 :42-53. (CBA-210)

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

3. Gu, L. et al. (2010). Stat5 promotes metastatic behavior ofhuman prostate cancer cells in vitro and in vivo. Endocr. Re-lat. Cancer 17 :481-493. (CBA-215)

CytoSelect Microfluidic Biochips


CytoSelect 8-Channel ECM Microfluidic Biochips Microscopy2 Chips CBA-003

10 Chips CBA-003-5

CytoSelect 8-Channel Endothelial Microfluidic Biochips Microscopy2 Chips CBA-004

10 Chips CBA-004-5

CytoSelect 8-Channel Microfluidic Biochips providean environment that closely mimics in vivo shearstresses, resulting in more physiologically relevantcell adhesion data. The Biochips are self-containedunits containing 8 channels with inlet/outlet ports atboth ends of each channel. After adding cell samples,a syringe pump set to the proper flow rate applies theappropriate shear stress to the channel.



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CELL-BASED A SSAYS Cell Migration / Invasion

1www.cellbiolabs.com [email protected]

Radius Cell Migration Assays (2D Gap Closure)


Radius 24-Well Cell Migration Assay Microscopy24 Assays CBA-125


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

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

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

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

Radius 96-Well Cell Migration Assay Microscopy96 Assays CBA-126


Radius 384-Well Cell Migration Assay384 Assays CBA-127

5 x 384 Assays CBA-127-5Microscopy

Radius Cell Migration Assays provide a unique al-ternative to the traditional Boyden Chamber migrationassay. Radius assays allow you to measure cell mi-gration at endpoint or in real time, and are ideal fortime course migration studies.

Radius Cell Migration Assays use a cell cultureplate containing a proprietary, carefully-defined bio-compatible hydrogel (Radius gel) spot centralizedat the bottom of each well. Cells seeded in the wellwill attach everywhere except on the Radius gel spot,creating a cell-free zone. Once cells attach, the Ra-dius gel is removed and migration of cells across thecell-free zone begins. The gel removal step allowssynchronization of a zero time point to facilitate well-to-well comparisons.

With Radius Cell Migration Assays, there are nocell culture inserts; so you dont need to worry aboutwhich pore size to choose for your cell type. Any ad-herent cell may be used in the assay.

Radius assays are supplied in 24-well, 96-well and384-well formats. In addition, the 24-well assays areprovided with your choice of coatings for proper cellattachment:

UncoatedCollagen I-coatedFibronectin-coated

Laminin-coatedECM Array with 6 wells of each of the above(uncoated, Collagen I, Fibronectin, Laminin); idealif you are unsure which ECM protein may providethe best cell attachment Ass ay Pri nci ple for the Rad ius Cell Migr ation Ass ays .


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CytoSelect Cell Migration AssaysChemotaxis

Product Name Pore Size Detection Size Catalog Number

CytoSelect 24-Well Cell Migration Assay

3 m Fluorometric 12 Assays CBA-103


8 mColorimetric 12 Assays CBA-100


12 mColorimetric 12 Assays CBA-107





CytoSelect 96-Well Cell Migration Assay

Migration o f Human Fibrosarcoma HT-1080 Cells. Cells wereseeded at 30,000 cells per well of a 24-well plate and allowed tomigrate toward 10% FBS for 4 hours. Migratory cells were stained(above) and quantified in a fluorescence plate reader (data notshown).

Fast Results : Visualize chemotaxis in less than6 hours with most cell typesFlexible : Bottoms of membrane inserts are un-coated to allow use with any chemoattractantHigher Through put : 96-well format available forfluorescence plate readers

CytoSelect Cell Migration Assays are ideal formeasuring chemotaxis. The kits utilize polycarbonatemembrane inserts in 24-well or 96-well plates. Insertsare available with 4 different pore sizes to accommo-date a variety of cell types.

Ass ay Pri nci pl e for the CytoSelec t Cell Migr ation Ass ay.

10% FBS0% FBS


Recent Product Citations 1. Tabata, C. et al. (2010). Novel clinical role of angiopoietin-1 in

malignant pleural mesothelioma. Eur. Respir. J. 36(5) :1099-1105.(CBA-100)

2. 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)

3. Igarashi, J. et al. (2009). Transforming growth factor-1 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 thetumor site of prostate cancer patients directs a selective loss ofimmunological tolerance to CCL2 that could be amplified in abeneficial 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 itsphysical association with nitric oxide synthase-3 in experimentalsepsis. J. Immunol. 183 :4055-4066. (CBA-104)

7. Christophi, G. et al. (2008). Modulation of macrophage infiltrationand inflammatory activity by the phosphatase SHP-1 in virus-induced demyelinating disease. J. Virol. 83 :522-539. (CBA-105)

8. Saher, H. et al. (2010). Red wine consumption improves in vitromigration of endothelial progenitor cells in young healthy individu-als. Am. J. Clinical Nutrition 92 :161-169. (CBA-106)



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CytoSelect Cell Migration AssaysHaptotaxis


CytoSelect 24-Well Cell Haptotaxis Assay, Collagen I-coatedColorimetric 12 Assays CBA-100-COL

Fluorometric 12 Assays CBA-101-COL

Colorimetric 12 Assays CBA-100-FN

Fluorometric 12 Assays CBA-101-FNCytoSelect 24-Well Cell Haptotaxis Assay, Fibronectin-coated

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

Haptotaxis describes the migration of cells toward agradient of immobilized extracellular matrix. TheCytoSelect Cell Haptotaxis Assays are ideal fordetermining the migratory properties of cells. Thekits utilize polycarbonate membrane inserts with an8 m pore size in a 24-well plate.

The undersides of the inserts are coated with eitherCollagen or Fibronectin. The 8 m pore size in themembrane inserts is ideal for epithelial cells, endo-thelial cells, fibroblasts, and other cells of similarsize. The membrane serves as a barrier that allowsdiscrimination of migratory cells from non-migratorycells.

Fast Results : Visualize cell haptotaxis inless than 6 hours with most cell typesConvenient : Membrane inserts pre-coatedon the underside with either Collagen I orFibronectinVersatile : Useful with a variety of cell typesincluding epithelial cells, endothelial cells,and fibroblasts*

*For leukocyte migration a 3 m 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 adhesionregulates vascular smooth muscle cell migration. J. Surg. Res.141:91-96. (CBA-100-COL)

Ass ay Pri nci ple for the CytoSelec t Cell Hapto taxi s As say.

BSA Collagen I Fibronectin

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CytoSelect Cell Migration AssaysTransmigration


CytoSelect Leukocyte Transmigration Assay Fluorometric 24 Assays CBA-212

Fluorometric 24 Assays CBA-216CytoSelect Tumor Transendothelial Migration Assay

Pore Size

3 m

8 m

Cancer cell transmigration, particularly extravasa-tion, is an important step in cancer metastasis. TheCytoSelect Cell Transmigration Assays provide arobust system for the quantitation of transmigrationsand interactions between endothelium and cancercells. 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. Xu, Z. et al. (2010). Role of pancreatic stellate cells in pancreatic

cancer metastasis. Am. J. Pathol . 177 :2585-2596. (CBA-216)

3. Yang, H. and H.E. Grossniklaus (2010). Constitutive overexpres-sion of pigment epithelium derived factor inhibition of ocularmelanoma growth and metastasis. Invest. Ophthalmol. Vis. Sci.51 :28-34. (CBA-216)

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The Leukocyte Adhesion and Transmigr ation Cascade.

Quantitation of Human Monocytic THP-1. LeukoTrackerlabeled THP-1 cells were titrated in 1X PBS, then lysed with 2X

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CytoSelect Cell Invasion Assays

Quantitative : Measure results in a colorimet-ric or fluorescence plate readerFlexible : Uniform protein matrix layer of yourchoice of basement membrane (from mousetumor cells), Collagen I, or Laminin IVersatile : Characterize both the invasive andmigratory properties of your cells with a CellMigration / Invasion Combo Kit (next page)

CytoSelect Cell Invasion As say Principl e.

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 onthe membrane bottom were stained (top and center) and quantifiedat OD 560nm after extraction (data not shown).

Tumor cell invasion into surrounding normal tissuecontributes to the morbidity of cancers. The CytoSe-lect Cell Invasion Assays use precoated inserts toassay invasive properties of tumor cells in 24-well or96-well plates. The coated layer serves to distinguishinvasive cells from non-invasive cells. Plates are pre-coated with either basement membrane matrix (fromEHS mouse sarcoma cells), Collagen I or Laminin I.

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Recent Product Citations 1. Zhang, L. et al. (2011). MicroRNA-1258 suppresses breast can-

cer brain metastasis by targeting heparanase. Cancer Res.71 :645-654. (CBA-110)

2. Coon, B. et al. (2010). The epsin family of endocytic adaptorspromotes fibrosarcoma migration and invasion. J. Biol. Chem.285 :33073-33081. (CBA-110)

3. Hirata, H. et al. (2010). Role of secreted Frizzled-related pro-tein3 in human renal cell carcinoma. Cancer Res. 70 :1896-1905.(CBA-110)

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

5. Eckstein, N. et al. (2009). Hyperactivation of the insulin-likegrowth factor receptor I signaling pathway is an essential eventfor 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 andenhances EMT stimulated by TGF through a PGE2-dependentmechanism. Carcinogenesis 29 :2227-2235. (CBA-112)

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



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CytoSelect Cell Invasion Assays, continued


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


CytoSelect 24-Well Cell Invasion Assay, Collagen IColorimetric 12 Assays CBA-110-COL

Fluorometric 12 Assays CBA-111-COL

Colorimetric 12 Assays CBA-110-LN

Fluorometric 12 Assays CBA-111-LN

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

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

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

CytoSelect 24-Well Cell Invasion Assay, Laminin I

CytoSelect Cell Migration / Invasion Assay Combo Kits

Effects of Cytoch alasin D on Invading Cells u sing the CytoSelect 24-well Cell Invasion Ass ay (CBA-110). HT-1080 and NIH3T3cells (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 2M 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 andinvasive properties of your cells. Each 24-well combo kit provides sufficient reagents to perform 12 migrationplus 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. Shin, S.Y. et al. (2010). TNFalpha-exposed bone marrow-derived mesenchymal stem cells promote locomotion of MDA-MB-231 breast

cancer cells through transcriptional activation of CXCR3 ligand chemokines. J. Biol. Chem. 285 :30731-30740. (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. Axlund, S.D. et al. (2010). HOXC8 inhibits androgen receptor signaling in human prostate cancer cells by inhibiting SRC-3 recruitement todirect androgen target genes. Mol. Cancer Res . 8:1643-1655. (CBA-106-C)

4. 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)

Product Name Pore Size Detection Size Catalog Number

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

Fluorometric 2 x 12 Assays CBA-101-C

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

8 m



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CytoSelect 24-Well Wound Healing / Cell Migration Assay

Highly Accurate : More consistent results well-to-well compared to homemade scratch assaysVersatile : Measure cell migration, cell prolifera-tion, and wound closure Inert Material : No residues from inserts to impedecell migration or proliferation

Wound healing assays are useful for studying tissuematrix remodeling, regulation of cytoskeletal struc-ture, and cell proliferation and migration rates of dif-ferent cells and culture conditions. Traditional woundhealing assays are performed by making a scratchacross a confluent cell monolayer to create an opengap, mimicking a wound. Such scratch assays,however, lack a consistently defined wound areaand result in high inter-sample variation.

Our CytoSelect 24-Well Wound Healing Assayprovides a more consistent method to measure cellmigration across a wound field gap in vitro . Pro-prietary treated inserts generate a consistently de-fined 0.9mm gap between the cells. Cells can thenbe 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 Assay24 Assays CBA-120

5 x 24 Assays CBA-120-5Microscopy

Wound Closur e of STO Cells. STO cells (mouse MEF) were cul-tured in the provided plate with inserts in place for 24 hours until amonolayer formed. Inserts were then removed to begin the assay.Cells were monitored at various time points and stained accordingto the assay protocol. CytoSelect 24-well Wound Healing Assay Princi ple.

0%

50%

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Percent Wound Closure


Recent Product Citation Tao, Y. et al. (2011). Treatment of breast cancer cells with DNAdemethylating agents leads to a release of Pol II stalling at geneswith DNA-hypermethylated regions upstream of TSS. Nucleic AcidRes . 10.1093/nar/gkr611.


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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 phagocytosisassays involve manually counting the engulfed sub-strates under a microscope. This process is tediousand time-consuming, can be somewhat inaccurate,and is not amenable to high throughput.

CytoSelect 96-Well Phagocytosis Assays are moreaccurate, high-throughput alternatives to the standardphagocytosis assay. The assays may be adapted foruse 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 A SSAYS Phagocytosis

Ass ay Pri nc ip le for t he Cyt oSelec t 96-Well Ph agocyt os is Ass ay (Zymosan).

Particle Engulfment with the CytoSelect 96-WellPhagocytosis As say (Zymosan).

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

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

2. Winnicka, B. et al. (2010). CD13 is dispensible for normal hema-

topoiesis and myeloid cell functions in the mouse. J. Leukoc.Biol. 88(2) :347-359. (CBA-220)

Ass ay Pri nci pl e for the CytoSelect 96-Well Phagocyt os is Ass ay (Red Bl ood Cell ).



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CELL-BASED A SSAYS Cell Viability / Death

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CytoSelect Anoikis Assays


CytoSelect 24-Well Anoikis Assay Colorimetric /Fluorometric 24 Assays CBA-080

CytoSelect 96-Well Anoikis Assay Colorimetric /Fluorometric 96 Assays CBA-081

Ano ikis of Hu man Fi brobl ast BJ-TERT Cell s. 50,000 cells/wellwere seeded in a control plate (left) and a Poly-HEMA coated plate(right) and cultured for 24 hours. Cells on the control plate werestained with Calcein AM. Cells on the Poly-HEMA coated platewere stained with EthD-1.

This assay allows you to quantify and monitor anoikisin cells using a Poly-HEMA precoated plate. Livecells 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 cytometryQuantitative : Measure live and dead cells on afluorescence plate reader; live cells may also bequantified 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 formonitoring cell viability via metabolic activity.

Live cells are detected with MTT (colorimetric detec-tion) or Calcein AM (fluorometric); dead cells aredetected with EthD-1 reagent (fluorometric). All 3detection 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 issuitable 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 wereseeded at 50,000 cells/well and allowed to culture for 24 hours.Cells were then treated with and without Saponin. All cells werethen stained with Calcein AM and EthD-1. Top : Cells withoutSaponin 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, orflow cytometry

Quantitative : Measure live and dead cells on afluorescence plate reader; live cells may also bequantified on a standard microplate reader



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Cellular Senescence Assays


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

96-Well Cellular Senescence Assay (SA -gal Activity) FluorometricPlate Reader 120 Assays CBA-231

Quantitative Cellular Senescence Assay (SA -gal) Fluorescence Micros-copy / Flow Cytometry 10 Assays CBA-232

Senescence Associated (SA) -galactosidase is a common biochemical marker of cellular senescence. Cellsexpressing such markers have been identified in vivo in tissues. SA -Gal catalyzes hydrolysis of X-gal, whichproduces a blue color in senescent cells.

Cell Contraction Assay (Collagen-Based)

CELL-BASED A SSAYS Senescence, Cell Contraction


Cell Contraction Assay Light Microscopy 24 Assays CBA-201

Collagen-Based Cell Contraction Assay Principl e.

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

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

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Recent Product Citation Schell, C. et al. (2010). 15-deoxy-delta12-14-prostaglandin-J2induces hypertrophy and loss of contractility in human testicularperitubular cells: implications for human male fertility. Endocrinol-ogy 151 :12571268. (CBA-201)

Our Cell Contraction Assay provides a simple systemto assess cell contractivity and to screen for cell con-traction mediators. The system uses a 3D collagenmatrix to measure changes in the collagen gel size.

An optional contraction inhibitor is provided.


The results may be seen using our SA -gal StainingKit or by flow cytometry using our Quantitative Cellu-lar Senescence Assay. For higher throughput, quan-tify cells on a fluorometric plate reader using our 96-Well Cellular Senescence Activity Assay.

Recent Product Citation

Malhotra, D. et al. (2010). Global mapping of binding sites for Nrf2identifies novel targets in cell survival response through ChIP-Seqprofiling and network analysis. Nucleic Acid Res . 10.1093/nar/gkq212. (CBA-231)


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Endothelial Tube Formation ( In Vitro Angiogenesis) Assay

HUVEC Tube Form ation 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 tumorcells. These cells proliferate to form new blood vessels.


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

CELL-BASED A SSAYS Angiogenesis, Autophagy

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Recent Product Citations 1. Kimura, T. et al. (2011). P2y5/LPA6 attenuates LPA1-mediated

VE-cadherin translocation and cell-cell dissociation throughG12/13 protein-Src-Rap1. Cardiovasc. Res. 10.1093/cvr/cvr154.

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

3. Weskamp, G. et al. (2010). Pathological neovascularization isreduced by inactivation of ADAM17 in endothelial cells but not inpericytes. Circ. Res. 106 :932-940 .

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

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

GFP-LC3 Expression Vectors

MAP LC3 is the most published autophagosomemarker protein. LC3 associates to the inner and outerlimiting membranes of the autophagosome. Twoforms of LC3 are visible by immunoblot: LC3I found inthe soluble fraction, and LC3II found in the mem-brane fraction. LC3II increases during autophagy.

Our GFP-LC3 expression vectors are available inthree formats: mammalian, lentiviral, and retroviralexpression. Each vector contains a GFP reportergene, and a GFP control plasmid is included at noadditional charge.


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

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

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

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

pCMV-GFP-LC3 Vector Map.


Recent Product Citation

Tu, S.P. et al. (2011). IFN-gamma inhibits gastric carcinogenesisby inducing epithelial cell autophagy and T-cell apoptosis. CancerRes. 71 :4247-4259. (CBA-401)


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STEM C ELL 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 thismay be accomplished by the introduction of key genes into somatic cells by transductionwith various viral vectors or transfection of plasmids.

Retroviral and lentiviral vectors appear to achieve among the highest levels of efficiency ofiPS cell generation. We offer an extensive collection of vectors for iPS cell reprogramming.

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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 atthe University of Tokyo.* Each vector contains one of 6 factors shown to help reprogram adult fibroblasts intoiPS cells. Both human and mouse genes are available individually or in sets. Separate retroviral vectors areavailable 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 cellplasmids. For additional information on these cell lines please see page 61 .

Target Name Vector Backbone Catalog Number

Oct-3/4 pMXs RTV-705

Sox2 pMXs RTV-706

c-Myc pMXs RTV-707Klf4 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 VectorsTarget Name Vector Backbone Catalog Number

Oct-3/4 pMXs RTV-701

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

p53 shRNA pRetro RTV-410

Human iPS Vectors


Platinum-E Retroviral Packaging Cell Line, Ecotropic >3 x 10 6 cells RV-101

Platinum-A Retroviral Packaging Cell Line, Amphotropic >3 x 10 6 cells RV-102

Platinum-GP Retroviral Packaging Cell Line, Pantropic >3 x 10 6 cells RV-103

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

Retroviral Packaging Cell Lines

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



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Induced Pluripotent Stem Cells

Lentiviral Polycistronic Vector for iPS Cell Generation

STEM C ELL RESEARCH

2


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

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

Our pLentG-KOSM Lentiviral Vector provides a con-venient way to generate iPS cells. The defined stemcells 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 transcriptionfactor ORF is followed by IRES-GFP as a reporter toverify viral transduction into your target cell. Efficien-cies of iPS generation are typically higher comparedto transduction of four separate viruses each contain-ing a single gene.

p53 shRNA has recently been shown to potentiallyincrease efficiency of iPS production. Our p53 shRNAlentiviral vector may be transduced along with thepLentG-KOSM vector.

More Efficient : Up to 10-fold higher efficiencycompared to multi-virus transduction, and 500-foldcompared to non-viral methodsReporter Convenience : Includes GFP reportergene to monitor lentiviral transduction

Open Reading Frame of pLentG-KOSM Lentiviral Vector.

Characterization o f iPS Cell Colo nies Generated from MEFsInfected with Lentivirus Cont aining the KOSM Fusion. Top : Staining of pluripotency markers in induced cell colonies at 200xmagnification. 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 Westernblot. Bottom : GFP fluorescence in MEF cells 3 days after infectionwith lentivirus containing KOSM fusion.

For efficient packaging of your KOSM lentivirus, please see our ViraSafe LentiviralPackaging Systems on pages 49-50 .



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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-305Platinum 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

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

Higher Viral Yields : Average titer 10 7 infec-tious units/mL with transient transfectionVersatile : 3 Packaging cell lines for use withnearly any target host species Optimized for Stem Cell Studies : Speciallydesigned expression systems for either ES/ECcells or hematopoietic stem cells

Ampho tro pi c Ecot ropic Pantr opic

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 asVSVG.

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 Platinu m Retroviral Expression Systems by HostSpecies.

Components of th e 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 plasmidtransfection. The Platinum Expression Systems in-clude one of our exclusive Platinum Packaging CellLines which already contain the gag and pol genes;the Ecotropic and Amphotropic cells also contain anenvelope protein. Simply clone your gene of interestinto the vector provided and transfect into the Plati-num cells. If you choose a Pantropic system, simplyco-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 avariety of cells, please see page 60 .

STEM C ELL RESEARCH Retroviral Expression Systems



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MEF Feeder Cells


MEF Feeder Cells 5 x 10 6 cells CBA-310

MEF Feeder Cells, Hygromycin-resistant 5 x 10 6 cells CBA-313

MEF Feeder Cells, Neomycin-resistant 5 x 10 6 cells CBA-311

MEF Feeder Cells, Puromycin-resistant 5 x 10 6 cells CBA-312

2

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

Feeder Cells STEM C ELL RESEARCH

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

The SNL 76/7 is an immortalized cell line derivedfrom mouse fibroblast STO cells which have beentransformed with murine LIF and neomycin resistancegenes.

Stem Cell FeedersLeukemia inhibitory factor (LIF) is useful for maintaining the undifferentiated state ofmouse 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 forboth 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 genefor better maintenance of undifferentiated state

Versatile : Useful for culture of human andmouse iPS cells and as a feeder for ES cellsPassage-Independent : Immortalized cell line


SNL Feeder Cells 3 x 10 6 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 pigmentsynthesis, cell proliferation, and epithelial morphology. Invest.Ophthalmol. Vis. Sci. 50 :5080-5088.

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

4. Lan, Z-J. et al. (2009). Extra-germ cell expression of mousenuclear 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 thatsupports long-term proliferation of stem cells. It hasbeen 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 typesincluding pluripotent ES cells, germ-line derived stemcells such as SPCs and MASCs, and primordial germcell-derived EG cells.


JK1 Feeder Cells 1 x 10 6 cells CBA-315

JK1 Cells Support Maintenance

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



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CytoSelect 96-Well Hematopoietic Colony Forming Cell Assay

Hematopoietic stem cells (HSCs), when cultured in asuitable semisolid matrix such as methylcellulosesupplemented with cytokines & nutrients, proliferateto form discrete cell clusters or colonies. Such HSCsor hematopoietic progenitors are known as colony-forming cells (CFCs).

In classic CFC assays, cells are cultured in a 35mmdish for 14-21 days so the colonies can reach a cer-tain size for manual counting, which can be tediousand subjective.

The CytoSelect 96-Well Hematopoietic ColonyForming Cell Assay provides a high-throughputmethod to quantify CFCs in just 7-10 days with nomanual 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 Fluorometric96 Assays CBA-320


Fast Results : 7-10 days vs. 2-3 weeksPlate Reader Convenience : Eliminates manualcountingEasier Reagent Handling : Methylcellulose mediacan be handled using a pipet instead of a syringe

Ass ay Pri nc ip le fo r th e CytoSelect 96-Well Hemat opoi eticColony Forming Cell Assay.

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

STEM C ELL RESEARCH Colony Formation Assays



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3

StemTAG 96-Well Stem Cell Colony Formation Assay


StemTAG 96-Well Stem Cell Colony Formation Assay Fluorometric96 Assays CBA-325


Our StemTAG 96-Well Stem Cell Colony Formation Assay provides a high-throughput method to quantifyES 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 dyeincluded 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 studyof tumor stem cells.

Fast Results : 7-10 days vs. 2-3 weeks using con-ventional methodsVersatile : Quantify cells using fluorescent dye,measure alkaline phosphatase activity, or recovercells for further analysisPlate Reader Convenience : No manual cellcounting required

Anc ho rage-Independent Gr owth o f Mouse ES-D3 Cells .Top : Phase Contrast. Bottom : Alkaline Phosphatase Staining.

StemTAG Stem Cell Colony Formation Assay Princip le.

Colony Formation Assays STEM C ELL RESEARCH



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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 RNAMurine Embryonic Stem Cell Line D3 50 g CBA-304

Total ProteinMurine Embryonic Stem Cell Line D3 500 g 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 celldifferentiation/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 andNCAM/Ectoderm), and two controls (GAPDH and -Actin). Primers are suitable for either end-point or real-time(quantitative) PCR.

STEM C ELL RESEARCH Alk Phos Assays, Primers, RNA/Protein

StemTAG Alkaline Phosphatase Kits

StemTAG Alkaline Phosph atase Staining Kit. Murine embry-onic stem cells (ES-D3) were maintained in an undifferentiatedstate with LIF. To induce differentiation, LIF was withdrawn overseveral days. Various differentiation events were observed: cellsbecame flattened and enlarged with reduced proliferation. On day5, cells were stained according to the assay protocol.


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

ICC & Fluorometric 2 x 100 Assays CBA-308

Colorimetric 100 Assays CBA-301


StemTAG Alkaline Phosphatase Staining Kit ICC 100 Assays CBA-300

StemTAG Alkaline Phosphatase Activity Assay Kit

The StemTAG Alkaline Phosphatase Staining and Activity Assay Kits monitor AP activity via both immu-nocytochemistry staining and a colorimetric activityassay. The staining and activity assay kits are alsosold separately.

Fast Results : Staining and Activity Assay proto-cols each take less than 1 hourVersatile : Useful for human ES, EG and EC cells,as well as mouse ES and EG cells

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

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

2. Izadyar, F. et al. (2008). Generation of multipotent cell lines froma distinct population of male germ line stem cells. Reproduction135 :771-784. (CBA-300)

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

4. Yue, Y. et al. (2008). A single intravenous injection of adeno-

associated virus serotype-9 leads to whole body skeletal muscletransduction in dogs. Mol. Ther. 16(12) :1944-1952. (CBA-301)5. Ghosh, A. et al (2007). Efficient whole-body transduction with

trans-splicing AAV vectors. Mol. Ther. 15(4) :750-755. (CBA-301)



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VIRAL EXPRESSION Viral Vector Overview

Recombinant Viral Gene DeliveryRecombinant 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. 35-40

Adenovirusp. 41-48

Lentivirus(HIV-1, FIV, SIV)

p. 49-55

Retrovirus(MMLV)p. 56-64

Gene Expression Transientor Stable TransientTransientor Stable Stable

Will Infect Dividing Cells Yes Yes Yes YesWill Infect Non-Dividing Cells Yes Yes Yes No

Integrates into Target Cell Genome No* No Yes Yes

Relative Viral Titer XXX XXXX XXX XX

Relative Transduction Efficiency XXX XXXX XXX XX

Immune Response in Target Cells Very Low High Low Moderate

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Typical Workflow for Viral Gene Delivery

Clone Geneof Interest

PackageVirus

MeasureTiter

Concentrate& Purify

InfectTarget Cell

ViralExpression

Plasmid

PackagingPlasmidsand Cells

Purification &Concentration

Kit

ViralTransduction

Reagents

ViralQuantitation

Kit


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

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


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VIRAL EXPRESSION AAV Expression

3

Adeno-Associated Virus Kits & Reagents

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

AAV Helper Free Systems

AAV Helper Free Systems are availablefor a variety of serotypes and kit formats.

Ordering information for AAV HelperFree Expression Systems, Packaging

Systems, Expression Vectors, andControl Vectors may be found on the

following pages.

Production of recombinant AAV requires certaingenes from the adenovirus genome, which meansthat an adenovirus usually needs to be present. The

AAV Helper Free System eliminates the need for ahelper adenovirus. Most of the required adenoviralgenes (E2A, E4 and VA RNA) are provided in apHelper plasmid, while the required E1 gene is pro-vided by the 293 packaging cells.

Safer : pHelper plasmid eliminates the need for ahelper virusFlexible : Packaging vectors and expression vec-tors available separately or as one complete sys-temControl Vectors : GFP, Cre or LacZ

Premade AAV ControlsPurification KitsQuantitation / Titer KitTransduction Kits

Helper Free Expression SystemsHelper Free Packaging SystemsExpression & Control VectorsViral Packaging Cell Line

Gene Delivery usi ng t he AAV Helper Free System.

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



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


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

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

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

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

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

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

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

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

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

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

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

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

AAV 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:

AAV Expression VectorpHelper PlasmidpRep-Cap Plasmid (serotype specific)GFP Control Vector

AAV Helper Free Expression Systems are availablefor the following serotypes:

Native serotypes 1-6 AAV-DJ, engineered by DNA family shuffling toform a hybrid capsid from 8 different native sero-types; provides significantly higher infectivityrates in vitro (see table below)

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

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

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

AAV-DJ/8

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

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

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

HepG2 Hu Liver 3 100 16.7 0.3 1.7 5 0.3 ND 1250 0.5Hep1A Ms Liver 20 100 0.2 1.0 0.1 1.0 0.2 0.0 400 0.1

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

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

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

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

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

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

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

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

Immature DC Hu Monocyte-derived DC 2500 100 ND ND 222 2857 ND ND 200 NDMature DC Hu Monocyte-derived DC 2222 100 ND ND 333 3333 ND ND 100 ND


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AAV Helper Free Complete Expression Systems (continued)


3www.cellbiolabs.com [email protected]


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









AAV-4 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER4 AAV-4 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER4
















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AAV Helper Free Packaging Systems

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


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




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

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



AAV Helper Free Expression VectorsProduct Name Size Catalog Number

pAAV-MCS Expression Vector 10 g VPK-410

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

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

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

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

pAAV-IRES-Bsd Expression Vector10 g VPK-419

pAAV-MCS Promoterless Expression Vector 10 g VPK-411

AAV Helper Free Control Plasmids


pAAV-GFP Control Vector 10 g AAV-400

pAAV-Cre Control Vector 10 g AAV-401

pAAV-LacZ Control Vector 10 g AAV-402


AAV Premade Control Viruses


AAV1-GFP Control Virus 50 L AAV-301





All AAV-GFP premade viruses are provided at a concentration of 1 x 1012

GC/mL.


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3

ViraBind AAV Purification Kits

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

High Purity : No contamination bands as seenon SDS gelFast Results : Obtain purified virus in about 3hours High Yields : Recovery rate >60%

Electrophoretic Profi le of Purifi ed 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 Kit2 Preps VPK-141

10 Preps VPK-141-510 x 15cm dishes

Purification Procedur e for the ViraBind AAV Purification Ki t.

AAV Packaging Cell Line


1 x 10 6 cells AAV-100293AAV Cell Line

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


Recent Product Citation Uchida, S. et al. (2010). Early li fe stress enhances behavioral

vulnerability to stress through the activation of REST4-mediatedgene transcription in the medial prefrontal cortex of rodents. J.Neurosci. 30 :15007-15018. (VPK-140)


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QuickTiter AAV Quantitation Kit

Product Name Capacity/Prep Size Catalog Number

QuickTiter AAV Quantitaiton Kit Fluorometric 20 Assays VPK-145

Fast Results : Obtain purified virus in less than 2hours High Sensitivit y : Limit of detection 1 x 10 9 GC/mLfrom unpurified supernatant or 5 x 10 10 GC/mL frompurified AAV

Traditional AAV Quantitation by dot blot can be tedi-ous, time consuming, and suffer from high inter-assayvariability. Our QuickTiter AAV Quantitation Kituses a proprietary technology to quantify AAV nucleicacid content of unpurified AAV-2 or AAV-DJ, or frompurified AAV of any serotype.

025

50

75

100

125

150

175

200

0 25 50 75 100 125 150

AAV DNA STD (ng)

R F U

0

5

10

15

20

0 2 4 6 8 10

AAV DNA STD (ng)

R F U

AAV-2 DNA Standard Cu rve. The QuickTiter AAV-2 DNA Standard was diluted as described in the assay protocol. Fluorescence wasmeasured 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-201ViraDuctin AAV Transduction Reagent

40

*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 cansignificantly increase the transduction efficiency of

AAV vectors in both dividing and non-dividing cells.Increases are greatest in non-dividing cells, but evencells in S-phase show a noticeable increase in trans-

duction efficiencies.

Higher Efficiencies : Significantly increase rate ofinfection 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. Infectionrates appear to be highest in S-phase cells, which can account for a very small fraction of a cell population.



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VIRAL EXPRESSION Adenoviral Expression

4

Adenoviral Expression Kits & Reagents

Recombinant adenoviruses are excellent tools for introducing genetic material into hostcells, 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 KitsQuantitation / Titer KitsTransduction Reagents

Viral Expression SystemsViral Packaging Cell LinePremade Recombinant Adenoviruses

RAPAd Adenoviral Expression Systems

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

Aden ovirus Pr oduction us ing th e RAPAd Aden oviral Exp ress ion Sys tem.

Adenoviruses are useful for delivering genes into awide variety of cell types, but traditional recombina-tion methods take several months and require tediousplaque purification. Recent technologies have short-ened the time required but still produce relatively highlevels of replication-competent plaques.

RAPAd Adenoviral Expression Systems producerecombinant adenovirus in about 2 weeks with asubstantial reduction in wild-type adenovirus. Thesystems use a backbone vector from which the 5ITR, packaging signal and E1 sequences have beenremoved. Additionally, serial amplification of the re-combinant adenovirus does not increase the level ofreplication-competent adenovirus.

Virtually No Wild-Type Virus : Backbone vector

engineered to produce


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42


Dont have time to make your own adenovirus? Areyou 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 10 9 viral particles per vial. Theyare provided as 50 l aliquots at a concentration of1 x 10 11 viral particles/mL in TBS with 10% glycerol.

Target Name Catalog NumberNull Control (No gene) ADV-001

-Galactosidase ADV-002

Cre ADV-005

Firefly Luciferase ADV-008

GFP ADV-004

SEAP (Secretory Alkaline Phosphatase) ADV-003

Controls and Reporter GenesRecent Product Citations 1. Matsushita, T. et al. (2009). FGFR3 promotes synchondosis

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

2. Ackerman, W. et al (2008). Nuclear Factor-kappa B regulatesinducible 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 hypertensiverats. Am. J. Physiol. Heart Circ. Physiol . 294 :H2871-H2878.(ADV-002)

4. Jones, S.W. et al. (2009). Mi togen-activated protein kinase-activated protein kinase (MK2) modulates key biological path-ways associated with OA disease pathology. Osteoarthritis andCartilage 17 :124-131. (ADV-004)

5. Perez-Moreno, M. et al. (2008). Loss of p120 catenin and linksto mitotic alterations, inflammation, and skin cancer. PNAS105 :15399-15404. (ADV-004)

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

Premade Recombinant Adenoviruses

Cancer/Tumor Antigens

Target Name Catalog Number

Carbonic Anhydrase 9 (CA9) ADV-602

Carcinoembryonic Antigen (CEA) ADV-604

NY-ESO-1 ADV-601

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

Angiogenesis


VEGF ADV-101

HIF-1 ADV-100

Recent Product Citations

1. Stoletov, K. et al. (2010). Visualizing extravasation dynamics ofmetastatic tumor cells. J. Cell Sci . 123 :2332-2341. (ADV-101)2. 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)

3. 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 293cell 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 forsuperior transfection and greater viral yields.

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

is not essential for the procoagulant activity of tissue factor orfor its de-encryption. Blood 115 :4273-4283. (AD-100)

2. Fang, S. et al (2007). Coordinated recruitment of histone me-

thyltransferase G9a and other chromatin modifying enzymes inSHP-mediated regulation of hepatic bile acid metabolism. Mol.Cell Biol . 27 :1407-1424. (AD-100)


293AD Cell Line 1 x 10 6 Cells AD-100



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4

Premade Recombinant Adenoviruses, continued

Act in Cyto skel eton Stai ni ng . 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 cytoskeletonwith Rhodamine-coupled Phalloidin.

Cytoskeleton Regulation / Small GTPase


Cdc42 ADV-152

Cdc42 L61 (Constitutively Active) ADV-154

Cdc42 N17 (Dominant Negative) ADV-153

PAK1 ADV-202

PAK1 (H83L, H86L) ADV-203

PAK1 (H83L, H86L, K299R) ADV-205

PAK1 (K299R) ADV-207

PAK1 (L107E, T423E) ADV-206PAK1 (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). TGF1 stimulates connective tissue

growth factor (CCN2/CTGF) expression in human gingivalfibroblasts through a RhoA-independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGF1-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 potentialof 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 cellmorphogenesis. J. Cell Sci. 122 :929-936. (ADV-150, ADV-153,

ADV-154)5. Cheng, Z.-J. et al. (2010). Co-regulation of caveolar and Cdc42

-dependent fluid phase endocytosis by phosphocaveolin-1. J.Biol. Chem . 285 :15119-15125. (ADV-153)

6. Perez-Moreno, M. et al. (2008). Loss of p120 catenin and linksto mitotic alterations, inflammation, and skin cancer. PNAS105 :15399-15404. (ADV-156)

7. 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)

8. Brantley-Sieders, D. et al. (2008). The receptor tyrosine kinaseEphA2 promotes mammary adenocarcinoma tumorigenesisand metastatic progression in mice by amplifying ErbB2 signal.J. Clin. Invest . 118 :64-78. (ADV-157)

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

10.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 & Transcription Regulation


p53 ADV-501

p53 (Temperature Sensitive Mutant) ADV-502

p68 RNA Helicase ADV-505


DCC AD

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