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table of contents ordering information support cell-based assays transformation adhesion migration invasion wound healing phagocytosis viability/death senescence contraction angiogenesis 4 5 stem cell research ips cell reprogramming retroviral expression systems colony formation assays alkaline phosphatase assays feeder cells 25 viral expression adenovirus adeno-associated virus aav lentivirus retrovirus 33 premade viruses expression systems purification kits titer kits transduction kits microrna analysis precursor clone collection mammalian expression vectors viral expression plasmids functional reporter system knockdown enhancer 65 2 phone 1-858-271-6500 usa toll-free 1-888-cbl-0505 fax 1-858-271-6514

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table of contents oxidative stress damage 75 protein oxidation/nitration lipid peroxidation dna/rna damage and repair reactive oxygen species ros assays antioxidant assays cell signaling protein biology small gtpase g-protein signaling kinase assays reporter cells and reagents epitope tags protein phosphorylation 97 metabolism research cholesterol assays apolipoprotein assays apolipoproteins and lipoproteins antibodies to apolipoproteins renal function assays 111 infectious disease research virus core antigen assays bacterial rapid detection kits 119 product index worldwide contacts www.cellbiolabs.com 123 130 info@cellbiolabs.com 3

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ordering information support placing an order within 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 american express® cards net 30 day terms may be offered upon credit approval worldwide technical support 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 benefit of their hands-on experience phone 1 858 271 6500 1 888 cbl 0505 us toll-free 1 858 271 6514 tech@cellbiolabs.com phone 1 858 271 6500 1 888 cbl 0505 toll-free 1 858 271 6514 orders@cellbiolabs.com www.cellbiolabs.com attn customer service 7758 arjons drive san diego ca 92126 fax e-mail fax e-mail online mail pricing current u.s prices are available online at www.cellbiolabs.com or you may request a separate price list by e-mail prices are subject 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 placing an order outside the u.s please see our worldwide distributors section on the inside back cover of this catalog we have a network of global distributors serving life science researchers in over 70 countries if you don t see your country listed please contact our u.s office kit components because our kits are qc tested by lot individual 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 sales@cellbiolabs.com 4 phone 1-858-271-6500 usa toll-free 1-888-cbl-0505 fax 1-858-271-6514

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cell-based assays colony formation assays tumor cell soft agar assays transformation of normal cells into neoplastic cells results in a population capable of proliferating independently of internal and external signals that normally restrain growth the soft agar colony formation assay has traditionally been used to monitor anchorageindependent 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 cytoselecttm 96-well cell transformation assay traditional soft agar colony formation our cytoselecttm 96-well cell transformation assay soft agar colony formation is suitable for measuring malignant transformation where no downstream analysis is required transformed cells cannot be recovered however no manual cell counting is required 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 recent product citations 1 inami y et al 2011 persistent activation of nrf2 through p62 in hepatocellular carcinoma cells j cell biol 1932 275-284 2 carnahan j et al 2010 selective and potent raf inhibitors paradoxically stimulate normal cell proliferation and tumor growth mol cancer ther 9:2399-2410 3 liu f et al 2010 epigenomic alterations and gene expression profiles in respiratory epithelia exposed to cigarette smoke condensate oncogene 29:3650-3664 4 iorns e et al 2010 the role of satb1 in breast cancer pathogenesis j natl cancer inst 10216 1284-1296 5 faoro l et al 2010 epha2 mutation in lung squamous cell carcinoma promotes increased cell survival cell invasion focal adhesions and mammalian target of rapamycin activation j biol chem 285:18575-18585 6 rubio r et al 2010 deficiency in p53 but not retinoblastoma induced the transformation of mesenchymal stem cells in vitro and initiates leiomyosarcoma in vivo cancer res 70:41854194 7 li h et al 2009 lysophosphatidic acid stimulates cell migration invasion and colony formation as well as tumorigenesis metastasis of mouse ovarian cancer in immunocompetent mice mol cancer ther 8:1692-1701 cell transformation assay principle product name cytoselecttm 96-well cell transformation assay soft agar colony formation detection fluorometric 5 plates cba-130-5 size 1 plate catalog number cba-130 fast results 6-8 days vs 21 days plate reader convenience eliminates manual counting versatile format designed for 96-well throughput but can be adapted for 48 24 12 or 6-well 6 phone 1-858-271-6500 usa toll-free 1-888-cbl-0505 fax 1-858-271-6514

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colony formation assays cell-based assays cytoselecttm 96-well cell transformation assays advanced soft agar with post-incubation cell recovery the cytoselecttm 96-well cell transformation assay cell recovery compatible provides a robust system for screening oncogenes and cell transformation inhibitors transformed cells may be recovered for further 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 throughput but can be adapted for 48 24 12 or 6-well 500 450 400 350 300 250 200 150 100 50 0 5000 2500 1250 625 313 0 hela nih3t3 rfu cells seeded/well easy fluorescence detection with the cytoselecttm cell transformation assay hela and nih3t3 cells were seeded at various concentrations and cultured for 6 days transformed colonies were quantified according to the assay protocol recent product citations 1 mathew b et al 2011 the novel role of the mu opioid receptor 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 protein3 in human renal cell carcinoma cancer res 70:1896-1905 cba-135 3 hirata h et al 2009 wnt antagonist gene dkk2 is epigenetically silenced and inhibits renal cancer progression through apoptotic and cell cycle pathways clin cancer res 15:56785687 cba-135 4 xie g et al 2009 acetylcholine-induced activation of m3 muscarinic receptors stimulates robust matrix metalloproteinase gene expression in human colon cancer cells am j physiol gastrointest liver physiol 296:g755-g763 cba-135 detection colorimetric cytoselecttm 96-well cell transformation assay cell recovery compatible fluorometric 5 plates 1 plate 5 plates 1 plate cytoselecttm 384-well cell transformation assay fluorometric 5 plates cba-145-5 cba-135-5 cba-140 cba-140-5 cba-145 size 1 plate catalog number cba-135 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 product name 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 www.cellbiolabs.com info@cellbiolabs.com 7

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cell-based assays colony formation assays cytoselecttm 96-well in vitro tumor sensitivity assay the cytoselecttm in vitro tumor sensitivity assay provides a stringent anchorage-independent model for chemosensitivity testing and possible anticancer drug screening the assay uses a soft agar matrix to promote the colony formation of neoplastic cells in about a week cells are quantified using a standard elisa plate reader fast results 6-8 days in vivo simulation resembles a threedimensional cell environment plate reader convenience eliminates manual counting recent product citations 1 itamochi h et al 2011 inhibiting the mtor pathway synergistically enhances cytotoxicity in ovarian cancer cells induced by etoposide through upregulation of c-jun clin cancer res 17:4742-4750 2 kang d.w et al 2010 phospholipase d1 drives a positive feedback loop to reinforce the wnt/ß-catenin/tcf signaling axis cancer res 70:4233-4242 tumor sensitivity assay principle 0.60 0.50 od 570nm 0.40 0.30 0.20 0.10 0.00 -11 -9 -7 -5 -3 log [5-fu m 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 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 according to the assay protocol ic50 value of 5-fluorouracil on hela cell anchorage-independent growth was determined to be 1 m product name cytoselecttm 96-well in vitro tumor sensitivity assay detection colorimetric size 96 assays 5 x 96 assays catalog number cba-150 cba-150-5 8 phone 1-858-271-6500 usa toll-free 1-888-cbl-0505 fax 1-858-271-6514

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colony formation assays cell-based assays cytoselecttm clonogenic tumor cell isolation kit clean separation of clonogenic tumor cells from normal cells is critical for proper analysis of disease state progression due to the heterogeneity of many tumors however isolation of homogenous tumor cell populations can be difficult the cytoselecttm 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 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 clonogenic tumor cell isolation procedure product name cytoselecttm clonogenic tumor cell isolation kit size 5 preps 25 preps catalog number cba-155 cba-155-5 www.cellbiolabs.com info@cellbiolabs.com 9

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cell-based assays cell adhesion cell adhesion assays cell adhesion is a complex mechanism involved in a variety of processes including cell migration/invasion embryogenesis wound healing and tissue remodeling cells can adhere to the ecm forming complexes with cytoskeletal components or to the endothelium our cytoselecttm cell adhesion assays quantify adhesion of cells using a microplate reader or fluorometer no manual cell counting is required cytoselecttm ecm cell adhesion assays the cytoselecttm 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 adherent cells attach while non-adherent cells are washed away adherent cells can be quantified on a standard plate reader or fluorometer quantitative measure results in a colorimetric or fluorescence plate reader flexible uniform substrate layer of your choice of collagen i collagen iv fibrinogen fibronectin or laminin or choose the ecm array which contains all 5 ecm proteins recent product citations 1 cervera a.m et al 2008 cells silenced for sdhb expression 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 studies of the optic nerve head astrocyte transcriptome from normal african americans and caucasian americans donors plos one 38 e2847 cba-060 bsa vitronectin laminin fibronectin collagen iv collagen i mda-231 ht-1080 hek293 cytoselecttm 48-well cell adhesion assay serum starved cells from three different cell lines were allowed to attach to the ecmcoated 48-well plate for 1 hr at 100,000 cells/well adherent cells were stained according to the assay protocol product name cytoselecttm 48-well cell adhesion assay ecm array contains one row each of collagen i collagen iv fibrinogen fibronectin and laminin cytoselecttm 48-well cell adhesion assay collagen i detection colorimetric fluorometric colorimetric fluorometric colorimetric size 48 assays 48 assays 48 assays 48 assays 48 assays 48 assays 48 assays 48 assays 48 assays 48 assays 48 assays 48 assays catalog number cba-070 cba-071 cba-052 cba-053 cba-060 cba-061 cba-058 cba-059 cba-050 cba-051 cba-056 cba-057 cytoselecttm 48-well cell adhesion assay collagen iv fluorometric colorimetric cytoselecttm 48-well cell adhesion assay fibrinogen fluorometric colorimetric cytoselecttm 48-well cell adhesion assay fibronectin fluorometric colorimetric cytoselecttm 48-well cell adhesion assay laminin fluorometric 10 phone 1-858-271-6500 usa toll-free 1-888-cbl-0505 fax 1-858-271-6514

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cell adhesion cell-based assays cytoselecttm leukocyte endothelium adhesion assays the cytoselecttm leukocyte endothelium adhesion assays provide a robust system for the quantitative determination of interactions between leukocytes and endothelium adherent cells can be quantified on a fluorescence plate reader recent product citations 1 xue j et al 2009 nfkb regulates thrombin-induced icam1 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 2 hernandez l et al 2010 activation of nf-kb signaling by inhibitor of nf-kb kinase ß increases aggressiveness of ovarian cancer cancer res 70:4005-4014 cba-215 3 gu l et al 2010 stat5 promotes metastatic behavior of human prostate cancer cells in vitro and in vivo endocr relat cancer 17:481-493 cba-215 140 120 100 rfu 80 60 40 20 0 25,000 50,000 thp-1 cells human monocytic thp-1 adhesion to huvec monolayer using the cytoselecttm leukocyte-endothelium adhesion assay huvec monolayer was treated with 1 µm pma for 12 hrs leukotrackertm 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 detection fluorometric fluorometric fluorometric size 96 assays 96 assays 96 assays catalog number cba-210 cba-211 cba-215 ctrl pma 100,000 cytoselecttm leukocyte-endothelium adhesion assay principle product name cytoselecttm leukocyte-endothelium adhesion assay cytoselecttm leukocyte-epithelium adhesion assay cytoselecttm tumor-endothelium adhesion assay cytoselecttm microfluidic biochips cytoselecttm 8-channel microfluidic biochips provide an environment that closely mimics in vivo shear stresses resulting in more physiologically relevant cell adhesion data the biochips are self-contained units containing 8 channels with inlet/outlet ports at both ends of each channel after adding cell samples a syringe pump set to the proper flow rate applies the appropriate shear stress to the channel product name cytoselecttm 8-channel ecm microfluidic biochips detection microscopy 10 chips 2 chips cytoselecttm 8-channel endothelial microfluidic biochips microscopy 10 chips cba-004-5 cba-003-5 cba-004 size 2 chips catalog number cba-003 www.cellbiolabs.com info@cellbiolabs.com 11

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cell-based assays cell migration invasion cell migration invasion assays cell migration and invasion are highly integrated multi-step processes and play important roles in the progression of various diseases including cancer atherosclerosis and arthritis our cell migration assays are provided in two formats 2-dimensional gap closure and boyden chamber each format has its own advantages and applications use the information below to help choose the best format for your cell migration experimental goals cell invasion assays are provided in the boyden chamber format cell migration format selection guide 2d gap closure assays p 13 type of analysis detection time detection method chemoattractant gradient relative sensitivity adaptability to automation cell compatibility qualitative or quantitative endpoint or real time microscopy no good good universal boyden chamber assays p 14-19 quantitative endpoint plate reader yes fair poor choose membrane pore size based on cell type/size example results using 2d gap closure assay example of boyden chamber assay principle 12 phone 1-858-271-6500 usa toll-free 1-888-cbl-0505 fax 1-858-271-6514

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cell migration invasion cell-based assays radiustm cell migration assays 2d gap closure radiustm cell migration assays provide a unique alternative to the traditional boyden chamber migration assay radius assays allow you to measure cell migration at endpoint or in real time and are ideal for time course migration studies radiustm cell migration assays use a cell culture plate containing a proprietary carefully-defined biocompatible hydrogel radiustm gel spot centralized at the bottom of each well cells seeded in the well will attach everywhere except on the radius gel spot creating a cell-free zone once cells attach the radius gel is removed and migration of cells across the cell-free zone begins the gel removal step allows synchronization of a zero time point to facilitate wellto-well comparisons with radiustm cell migration assays there are no cell culture inserts so you don t need to worry about which pore size to choose for your cell type any adherent cell may be used in the assay radius assays are supplied in 24-well 96-well and 384-well formats in addition the 24-well assays are provided with your choice of coatings for proper cell attachment uncoated collagen i-coated fibronectin-coated laminin-coated ecm array with 6 wells of each of the above uncoated collagen i fibronectin laminin ideal if you are unsure which ecm protein may provide the best cell attachment product name radiustm 24-well cell migration assay radiustm 24-well cell migration assay collagen i coated radiustm 24-well cell migration assay fibronectin coated radiustm 24-well cell migration assay laminin coated radiustm 24-well cell migration assay ecm array coated radiustm 96-well cell migration assay assay principle for the radiustm cell migration assays detection microscopy 5 x 24 assays microscopy microscopy microscopy microscopy microscopy 5 x 96 assays 384 assays cba-126-5 cba-127 cba-127-5 24 assays 24 assays 24 assays 24 assays 96 assays cba-125-5 cba-125-col cba-125-fn cba-125-ln cba-125-ecm cba-126 size 24 assays catalog number cba-125 radiustm 384-well cell migration assay microscopy 5 x 384 assays www.cellbiolabs.com info@cellbiolabs.com 13

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cell-based assays cell migration invasion cytoselecttm cell migration and invasion assays boyden chamber the boyden chamber has been extensively used and widely published as a tool for measuring cell migration and cell invasion in vitro our cytoselecttm cell migration and invasion assays use this well-cited method to quantify cell migration and invasion with no manual cell counting required migratory or invasive cells are quantified using a colorimetric or fluorometric plate reader cell migration may take on various forms and behaviors depending on the type and location of cells such subclasses of cell migration include chemotaxis haptotaxis and transmigration use the chart below to compare the various subclasses of cell migration as well as cell invasion which will help you choose the assay best suited to your experimental goals typical well setup for boyden chamber assay boyden chamber assay selection guide assay definition cell types neutrophils leukocytes lymphocytes monocytes macrophages fibroblasts endothelial cells epithelial cells tumor cells pore size 3 µm insert coating none assay formats 24-well 96-well 24-well 96-well 5 µm none chemotaxis p 15 migration of cells toward a chemoattractant chemical signal in the cell s surrounding environment 8 µm none 24-well 96-well astrocytes 12 µm slow-moving cells migration of cells along a gradient of cellular adhesion sites or extracellular matrixbound chemoattractants fibroblasts endothelial cells epithelial cells leukocytes tumor cells none collagen i bottom 24-well 24-well 24-well 24-well 24-well 24-well 96-well 24-well 96-well 24-well 96-well haptotaxis p 16 8 µm fibronectin bottom 3 µm 8 µm none none ecm matrix top 8 µm collagen i top laminin i top migration of cells through the transmigration vascular endothelium toward a p 17 chemoattractant movement of cells through the 3d extracellular matrix into neighboring tissues includes ecm degradation and proteolysis invasion p 18-19 fibroblasts endothelial cells epithelial cells tumor cells 14 phone 1-858-271-6500 usa toll-free 1-888-cbl-0505 fax 1-858-271-6514

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cell migration invasion cell-based assays cytoselecttm cell migration assays chemotaxis cytoselecttm 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 accommodate a variety of cell types fast results visualize chemotaxis in less than 6 hours with most cell types flexible bottoms of membrane inserts are uncoated to allow use with any chemoattractant higher throughput 96-well format available for fluorescence plate readers 0 fbs 10 fbs 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 recent product citations 1 tabata c et al 2010 novel clinical role of angiopoietin-1 in malignant pleural mesothelioma eur respir j 365 1099-1105 cba-100 2 awasthi n et al 2009 endothelial monocyte activating polypeptide ii interferes with vegf-induced proangiogenic signaling laboratory investigation 891 38-46 cba-100 3 igarashi j et al 2009 transforming growth factor-ß1 downregulates caveolin-1 expression and enhances sphingosine 1phosphate 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 mediate term and preterm labor j immunol 181:1470-1479 cba102 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 christophi g et al 2008 modulation of macrophage infiltration and inflammatory activity by the phosphatase shp-1 in virusinduced demyelinating disease j virol 83:522-539 cba-105 8 saher h et al 2010 red wine consumption improves in vitro migration of endothelial progenitor cells in young healthy individuals am j clinical nutrition 92:161-169 cba-106 detection fluorometric fluorometric colorimetric size 12 assays 12 assays 12 assays 12 assays 12 assays 12 assays 96 assays 96 assays 96 assays catalog number cba-103 cba-102 cba-100 cba-101 cba-107 cba-108 cba-104 cba-105 cba-106 assay principle for the cytoselecttm cell migration assay product name pore size 3 µm 5 µm cytoselecttm 24-well cell migration assay 8 µm fluorometric colorimetric 12 µm fluorometric 3 µm cytoselecttm 96-well cell migration assay 5 µm 8 µm fluorometric fluorometric fluorometric www.cellbiolabs.com info@cellbiolabs.com 15

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