IL-10 MIP- M-CSF IL-2 G-CSF. Bio-Plex Multiplex System. IL-12p40 IL-4 MIP-2. IFN-γ. TGF-β3 IL-6. Multiplex System IL-5

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1 IL-β Multiplex System IL- TGF-β IL-6 M-CSF GLP- TGF-β ICAM- TRAIL TGF-β TGF-β CTACK MIF TGF-β TGF-β TGF-β MIF PAI- TGF-β HGF TGF-β TGF-β IL- IL-4 TGF-β IFN-Υ TGF-β TGF-β Bio-Plex Multiplex System TGF-β IL-5 TGF-β TGF-β TGF-β The Leader i Multiplex Immuoassay Aalysis IL- TGF-β IL-p7 TGF-β IL- TGF-β TGF-β IL- TGF-β TGF-β TGF-β G-CSF TGF-β TGF-β TGF-β IL-7 TGF-β MIP- MIP- IL- IFN-γ IL-p4 T

2 The Bio-Plex Multiplex System The Bio-Plex System is a flexible, easy-to-use microplate-based multiplex immuoassay system. It eables you to quatify up to differet aalytes i a sigle sample, givig you the power to uderstad complex relatioships amog proteis i ormal ad disease states. Multiplex aalysis with the Bio-Plex System provides more iformatio with less sample. The Bio-Plex System comprises all products for a complete workflow from sample prep reagets ad diluets to sophisticated data aalysis for usurpassed sesitivity, precisio, ad accuracy. Bio-Plex Multiplex Reader With the Bio-Plex System, you ca multiplex: Immuoassays Receptor-ligad assays Nucleic acid hybridizatio assays Ezyme assays Bio-Plex D Multiplex Reader Bio-Plex Pro Wash Statio Bio-Plex Pro Assays BIO-PLEX IMMUNOASSAY WORKFLOW From Multiplex Assay to Results i as Little as Hours Bio-Plex Assays are desiged usig microscopic beads, each with a differet color code (spectral address) to permit discrimiatio of idividual assays. Beads are dyed with differet ratios of two fluorophores ad are thus classified ito uique bead regios (xmap techology).. Coupled Beads. Samples A Bio-Plex Assay uses a selectio of beads with differet spectral addresses, each coupled to atibodies agaist a differet target. Sample types ragig from serum, urie, milk, ad plasma to cell lysates with a Bio-Plex Cell Lysis (prepared Kit) ca be used with various Bio-Plex Assays. Add coupled beads to wells of a 96-well plate Add samples to wells cotaiig coupled beads Wash Icubate mi mi Wash mi mi mi 4 mi 8 mi 7 mi 7 mi 8 mi 4 mi 8 mi 4 mi 4 mi 4 mi 7 mi mi 4 mi 8 mi 7 mi 7 mi 8 mi 4 mi 4 mi mi 4 mi 7 mi 4 mi 4 mi 4 mi 8 mi

3 A Multitude of Choices Bio-Plex Assay Fider Visit bio-rad.com/fidmyassay to browse available kits by aalyte ame, compatible species, ad research area. Bio-Rad offers a cotiually growig lie of assays ad associated products for extracellular ad itracellular aalysis. The assays are relevat to researchers studyig everythig from cacer to cardiovascular disease ad available for use with multiple species ad sample types. Go to bio-rad.com/bioplex for the most up-to-date target iformatio. Fid the right Bio-Plex Assay with oly a few clicks. Cytokies, chemokies, ad growth factors Browse the etire catalog of targets alphabetically Filter by species ad research area Multiplex Assays i a Variety of Pael Cofiguratios for your customized search results to facilitate orderig Cotact a Bio-Rad represetative who ca talk to you about your optios Isotypig TGF-b Acute phase The Th7 pathway MMP ad TIMP Bio-Plex Assay Builder Cell sigalig Iflammatio Desig your ow custom assays from our selectio of hudreds of aalytes. Go to bio-rad.com/assaybuilder to lear more. Cacer Diabetes Kidey toxicity ad kidey damage Develop Your Ow Assay Develop uique bead-based multiplex assays with Bio-Plex Pro COOH Beads. Bio-Plex Pro COOH Beads are carboxy-coated beads ready to be covaletly coupled to the protei or ucleic acid of your choice. 4. Reporter. Detectio Atibodies Bioti-labeled detectio atibodies are specific for secodary epitopes o each target. Add detectio atibodies to wells Wash mi 4 mi 7 mi 8 mi 7 mi 7 mi 8 mi 8 mi Resusped i assay buffer 4 mi 8 mi 8 mi Dyed beads are idetified 4 mi 4 mi by their iteral fluorescet sigature (bead regio) 7 mi 7 mi 7 mi = segfr (. Bulleti pg/ml) 8 mi 8 mi Multiplex data are reported simultaeously 8 mi Level of target boud to beads is idicated by itesity of reporter sigal 7 mi 4 mi 4 mi 4 mi 8 mi 4 mi 4 mi Rise Icubate mi 4 mi 5. Assay 4 mi 4 mi 7 mi 7 mi Bio-Rad Laboratories, Ic. 4 mi 4 mi 4 mi mi mi Icubate mi mi mi Fluorescetly labeled streptavidi The Bio-Plex Reader first classifies reporter bids beads by spectral addresses ad mi mi mi to the bioti-labeled 4 mi 4 mi 4 mi detectio atibodies. the quatifies the associated reporter sigal itesity. Add reporter to wells mi = svegfr (5. pg/ml) = sher- (.4 pg/ml) = IL-6Ra (.7 pg/ml)

4 Bio-Plex Software is the ideal solutio to reduce errors, save time, ad get up to speed fast. Reduce errors Save time Get up to speed fast Importig results directly ito a database from Bio-Plex Maager data files drastically reduces the eed to audit results ad elimiates mistakes itroduced by copy-adpaste errors Editig aalysis optios results i istataeous updates to all the outputs ad reduces the time spet creatig graphs ad performig aalysis Automatig calculatios based o the most commo comparisos used by biological researchers elimiates time spet orgaizig data Applyig a truly itegrated solutio simplifies ad speeds up data aalysis Truly user-friedly software removes traiig burde, esures istrumet reliability, ad elimiates user error BIO-PLEX SOFTWARE Bio-Plex Maager Software is the ideal tool for reviewig ru data ad optimizig results from ay xmap istrumet. Bio-Plex Data Pro Software combies multiple istrumet rus to efficietly uderstad what your multiplex data say about your biological system. Try Bio-Plex Data Pro free for days Visit bio-rad.com/ad/datapro to dowload it.

5 The Power of Bio-Plex Assays Bio-Plex Multiplex Assays deliver more data i less time with less sample ad with fewer maual maipulatios. The examples to the right show a compariso betwee a Bio-Plex Assay ad covetioal methods. I the top pael, a Bio-Plex Phosphoprotei 5-Plex Assay is compared with 5 wester blots. O the bottom pael, a Bio-Plex Huma Cytokie 7-Plex Assay is compared with 7 ELISA assays. Prove Results The Bio-Plex System is the most widely cited multiplex assay platform i life sciece research today. Applicatio areas iclude iflammatio, autoimmue disease, asthma, diabetes/obesity, cacer, eurological disorders, cardiovascular disease, drug discovery, ad vaccie developmet. Ope Platform Bio-Plex Pro Magetic Assays cofigured o MagPlex Beads are compatible with all xmap life sciece istrumets supplied by ay Lumiex parter. Choose from a broad selectio of precofigured ad custom assay paels. Aalysis of 5 phosphoproteis i samples. Wester Blot Bio-Plex System Number of phosphoproteis 5 5 Number of samples Total data poits 6 6 Number of gels or 96-well plates 5 Data poits per gel or plate 6 Total time required ~8 hr + oe hr + oe overight icubatio overight icubatio Sample volume 5 µl 5 µl Dyamic rage Limited total protei.4 µg/well* * The rage of total lysate protei i which the aalyte ca be detected. Aalysis of 7 cytokies i 8 samples. ELISA Bio-Plex System Number of cytokies 7 7 Number of samples 8 8 Total data poits,6,6 Number of 96-well plates 7 Data poits per plate 8,6 Total time required >6 hr hr Sample volume Serum or plasma, Serum or plasma, > ml*.5 µl Cell culture superatat, Cell culture superatat, > ml* 5 µl * Based o 5 µl/well of sample. Bio-Plex Maager Software supports all Lumiex ad istrumets. I additio, it supports both magetic ad omagetic beads ad ca aalyze data from ay xmap platform. Kits are available to ru Bio-Plex Maager o Lumiex Assay Readers. For more iformatio, cotact your local Bio-Rad sales represetative. Lumiex, MagPlex, ad xmap are trademarks of the Lumiex Corporatio. The Bio-Plex Suspesio Array System icludes fluorescetly labeled microspheres ad istrumetatio licesed to Bio-Rad Laboratories, Ic. by the Lumiex Corporatio. CST Atibodies developed ad validated for Bio-Plex Phosphoprotei ad Total Target Assays. IL- TGF-β IL-α M-CSF IL-6 ICAM- TGF-β IL-7 TRAIL TGF-β TGF-β CTACK MIF TGF-β TGF-β

6 Bio-Rad Laboratories, Ic. Life Sciece Group Web site bio-rad.com USA Australia Austria Belgium () 7 5 Brazil Caada Chia Czech Republic Demark Filad Frace Germay Hog Kog Hugary Idia Israel Italy 9 69 Japa Korea Mexico The Netherlads () New Zealad Norway Polad Portugal Russia Sigapore South Africa 7 () Spai Swede Switzerlad Taiwa Thailad Uited Arab Emirates Uited Kigdom Bulleti 545 Ver E US/EG Sig 5

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