Multiplex Suspension Arrays NF-k p65 EGFR Smad2 TGF- c-jun Akt IL-17A/F Stat1 IL-12p70 IGF-IR MEK1 Bio-Plex

Transcription

1 Multiplex Suspension Arrays NF-kβ p65 TGF-β1 c-jun Smad2 EGF IL-17A/F Akt IGF-IR Stat1 MEK1 IL-12p70 Bio-Plex Multiplex Immunoassays Analyte Guide Summer 2014

2 2014 Bio-Rad Laboratories, Inc.

3 The Power of Bio-Plex Assays 2 Bio-Plex Assay Configurations 4 Bio-Plex Comprehensive Analyte Tables 6 Circulating Magnetic Assays 6 Magnetic Cell Signaling Assays 14 Human Assays 18 Cytokine Group l 18 Cytokine Group ll 19 Th17 Cytokine 20 Chemokines 21 TGF-b 23 Cancer Biomarkers 24 Apoptosis 26 Diabetes 27 Metabolic and Hormone New 28 MMPs and TIMPs New 30 Kidney Toxicity 31 Isotyping 32 Acute Phase 33 Mouse Assays 34 Cytokine Group l 34 Cytokine Group ll 35 Cytokine Group lll 35 TGF-b 35 Diabetes 36 Rat Assays 37 Cytokine 37 TGF-b 37 Diabetes 38 Kidney Toxicity 39 Canine Assays 40 Kidney Toxicity 40 Bulletin

4 The Power of Bio-Plex Assays Bio-Plex Multiplex Assays deliver more data in less time with less sample and with fewer manual manipulations. The examples on the next page show a comparison between a Bio-Plex assay and conventional methods. In the top panel, a Bio-Plex Phosphoprotein 5-Plex Assay is compared with five western blots. In the bottom panel, a Bio-Plex Human Cytokine 27-Plex Assay is compared with 27 ELISA assays. Proven Results The Bio-Plex System is the most widely cited multiplex assay platform in life science research today. Application areas include inflammation, autoimmune disease, asthma, diabetes/obesity, cancer, neurological disorders, cardiovascular disease, drug discovery, and vaccine development. Open Platform Bio-Plex Pro Magnetic Assays configured on MagPlex Beads are compatible with all xmap life science instruments supplied by any Luminex partner. Choose from a broad selection of preconfigured and custom assay panels. Bio-Plex Manager Software supports all Luminex 100 and 200 Instruments. In addition, it supports both magnetic and nonmagnetic beads and can analyze data from any xmap platform. Kits are available to run Bio-Plex Manager on Luminex Assay Readers. For more information, contact your local Bio-Rad sales representative Bio-Rad Laboratories, Inc. 2

5 Analysis of five phosphoproteins in 12 samples. Western Blot Bio-Plex System Number of phosphoproteins 5 5 Number of samples Total data points Number of gels or 96-well plates 5 1 Data points per gel or plate Total time required ~8 hr + one 2 hr + one overnight incubation overnight incubation Sample volume 125 µl 50 µl Dynamic range Limited total protein µg/well* * The range of total lysate protein in which the analyte can be detected. Analysis of 27 cytokines in 80 samples. TGF-β1 c-jun NF-kβ p65 Smad2 EGFR ELISA Bio-Plex System Number of cytokines Number of samples Total data points 2,160 2,160 IL-17A/F Akt Number of 96-well plates 27 1 Data points per plate 80 2,160 Total time required >60 hr 3 hr Sample volume Serum or plasma, Serum or plasma, >1 ml* 12.5 µl Cell culture supernatant, Cell culture supernatant, >1 ml* 50 µl * Based on 50 µl/well of sample. IGF-IR Stat1 PDGFR-α MEK1 TGF-β1 IL-12p70 c GSK-3α/β JNK c-jun Bulletin

6 Bio-Plex Assay Configurations A Multitude of Choices Bio-Rad offers a continually growing line of assays and associated products for extracellular and intracellular analysis. The assays are relevant to researchers studying everything from cancer to cardiovascular disease and available for use with multiple species and sample types. For the most up-to-date target information, go to bio-rad.com/bioplex. Bio-Plex Assay Builder Design your own custom assays from our selection of hundreds of analytes in four easy steps. With the Assay Builder you can: n Get a price quote n Place your order online n Look up an existing assay by catalog number Go to bio-rad.com/assaybuilder x-plex Assay Express Assay Develop Your Own Assay Develop unique bead-based multiplex assays with Bio-Plex COOH beads. Bio-Plex COOH beads are carboxyl-coated beads ready to be covalently coupled to the protein or nucleic acid of your choice. n Couple antibodies, antigens, receptors, ligands, enzymes, or nucleic acids n Couple protein (~6 150 kd) with 1 ml of beads in <5 hr for use in up to 20 microplates n The Bio-Plex Amine Coupling Kit is available for protein and antibody coupling We Mix You Mix 2014 Bio-Rad Laboratories, Inc. 4

7 Over 50% of multiplex assays worldwide are run using Bio-Plex Software. Try Bio-Plex Manager and Bio-Plex Data Pro Software and find out why. Bulletin

8 Bio-Plex Comprehensive Analyte Tables Bio-Plex Circulating Magnetic Assays. Target Human (p 18 33) Mouse (p 34 36) Rat (p 37 39) Non-Human Primate (p 27) Canine (p 40) 6Ckine / CCL21 Chemokine 40-plex (p 21) AAT Metabolic Panel 3 (p 28) Adiponectin Diabetes (p 27) Diabetes (p 36) Diabetes (p 27) Adipsin Diabetes 2-plex (p 27) Diabetes (p 27) AGP-1 Metabolic Panel 3 (p 28) Albumin Kidney Toxicity 2 6-plex (p 31) Kidney Toxicity (p 39) Kidney Toxicity (p 40) Angiopoietin-2 Cancer 2 18-plex (p 25) ANGPTL3 Metabolic Panel 4 (p 28) Bad Apoptosis 2 (p 26) Bak Apoptosis 1 (p 26) Bax Apoptosis 1 (p 26) Bax / Bcl-2 dimer Apoptosis 2 (p 26) BCA-1 / CXCL13 Chemokine 40-plex (p 21) Bcl-xL Apoptosis 2 (p 26) Bcl-xL / Bak dimer Apoptosis 3 (p 26) Bim Apoptsis 2 (p 26) B2M Kidney Toxicity 2 6-plex (p 31) Kidney Toxicity 2 4-plex (p 39) Calbindin Kidney Toxicity 1 6-plex (p 31) Kidney Toxicity 2 4-plex (p 39) Caspase-3, active Apoptosis 3 (p 26) CD40L Cancer 2 18-plex (p 25) Group III Cytokine Th17 (p 35) Chemerin Metabolic Panel 4 (p 28) Clusterin Kidney Toxicity 1 6-plex (p 31) Kidney Toxicity 1 5-plex (p 39) Kidney Toxicity 1 4-plex (p 40) Cortisol Metabolic Panel 1 (p 28) C-Peptide Diabetes 10-plex (p 27) Diabetes 10-plex (p 27) Metabolic Panel 1 (p 28) CRP Acute Phase 4-plex (p 33) CTACK / CCL27 Group II Cytokine 21-plex (p 19) Chemokine 40-plex (p 21) Cystatin C Kidney Toxicity 2 6-plex (p 31) Kidney Toxicity 2 4-plex (p 39) DPP4 Metabolic Panel 4 (p 28) EGF Cancer 2 18-plex (p 25) ENA-78 / CXCL Chemokine 40-plex (p 21) continues 2014 Bio-Rad Laboratories, Inc. 6

9 . Target Human (p 18 33) Mouse (p 34 36) Rat (p 37 39) Non-Human Primate (p 27) Canine (p 40) segfr Cancer 1 16-plex (p 24) Endoglin Cancer 2 18-plex (p 25) Eotaxin / CCL11 Group l Cytokine 27-plex (p 18) Group I Cytokine 23-plex (p 34) Chemokine 40-plex (p 21) Eotaxin-2 / CCL24 Chemokine 40-plex (p 21) Eotaxin-3 / CCL26 Chemokine 40-plex (p 21) EPO Cytokine 24-plex (p 37) sfasl Cancer 2 18-plex (p 25) Ferritin Acute Phase 5-plex (p 33) FGF-21 Metabolic Panel 2 (p 28) FGF-23 Metabolic Panel 2 (p 28) FGF-basic Cancer 1 16-plex (p 24) Group II Cytokine 9-plex (p 35) Group I Cytokine 27-plex (p 18) Fibrinogen Acute Phase 5-plex (p 33) Follistatin Cancer 1 16-plex (p 24) Fractalkine / CX3CL1 Chemokine 40-plex (p 21) FSH Hormone Panel 1 (p 28) Galectin-3 Metabolic Panel 2 (p 28) GCP-2 / CXCL6 Chemokine 40-plex (p 21) G-CSF Cancer 1 16-plex (p 24) Group I Cytokine Cytokine 24-plex (p 37) Group I Cytokine 23-plex (p 34) 17-plex/27-plex (p 18) GH Hormone Panel 1 (p 28) Ghrelin Diabetes 10-plex (p 27) Diabetes 8-plex (p 36) Diabetes (p 38) Diabetes 10-plex (p 27) GIP Diabetes 10-plex (p 27) Diabetes 8-plex (p 36) Diabetes 10-plex (p 27) GLP-1 Diabetes 10-plex (p 27) Diabetes 8-plex (p 36) Diabetes (p 38) Diabetes 10-plex (p 27) Glucagon Diabetes 10-plex (p 27) Diabetes 8-plex (p 36) Diabetes (p 38) Diabetes 10-plex (p 27) GM-CSF Group I Cytokine 8-plex/ Group I Cytokine Cytokine 24-plex/ 17-plex/27-plex (p 18) 8-plex/23-plex/[Th1/Th2] (p 34) [Th1/Th2] (p 37) Chemokine 40-plex (p 21) Gro-a / CXCL1 Group II Cytokine 21-plex (p 19) Chemokine 40-plex (p 21) Gro-b / CXCL2 Chemokine 40-plex (p 21) continues Bulletin

10 . Target Human (p 18 33) Mouse (p 34 36) Rat (p 37 39) Non-Human Primate (p 27) Canine (p 40) GST-π Kidney Toxicity 1 6-plex (p 31) Haptoglobin Acute Phase 4-plex (p 33) HB-EGF Cancer 2 18-plex (p 25) Hemopexin Metabolic Panel 3 (p 28) sher-2 / neu Cancer 1 16-plex (p 24) HGF Cancer 1 16-plex (p 24) Group II Cytokine 21-plex (p 19) I-309 / CCL1 Chemokine 40-plex (p 21) ICAM-1 Group II Cytokine singleplex (p 19) Group III singleplex (p 35) IFN-a2 Group II Cytokine 21-plex (p 19) IFN-g Group l Cytokine 8-plex/ Group I Cytokine 8-plex/ Cytokine Th1/Th2 (p 37) 17-plex/27-plex/[Th1/Th2] (p 18) 23-plex/[Th1/Th2]/Th17 (p 34) Chemokine 40-plex (p 21) IgA Isotyping 6-plex (p 32) IgE Isotyping singleplex (p 32) IGF-1 IGF Panel (p 28) IGF-2 IGF Panel (p 28) IGFBP-1 Cancer 2 18-plex (p 25) IGFBP Panel (p 28) IGFBP-2 IGFBP Panel (p 28) IGFBP-3 IGFBP Panel (p 28) IGFBP-4 IGFBP Panel (p 28) IGFBP-5 IGFBP Panel (p 28) IGFBP-6 IGFBP Panel (p 28) IGFBP-7 IGFBP Panel (p 28) IgG 1 Isotyping 6-plex (p 32) IgG 2 Isotyping 6-plex (p 32) IgG 3 Isotyping 6-plex (p 32) IgG 4 Isotyping 6-plex (p 32) IgG-total Isotyping singleplex (p 32) IgM Isotyping 6-plex (p 32) continues 2014 Bio-Rad Laboratories, Inc. 8

11 . Target Human (p 18 33) Mouse (p 34 36) Rat (p 37 39) Non-Human Primate (p 27) Canine (p 40) IL-1a Group ll Cytokine 21-plex (p 19) Group I Cytokine 23-plex (p 34) Cytokine Th1/Th2 (p 37) IL-1b Group I Cytokine Group I Cytokine Cytokine Th1/Th2 (p 37) 17-plex/27-plex (p 18) 8-plex/23-plex (p 34) Th17 Cytokine 15-plex (p 20) Chemokine 40-plex (p 21) IL-1Ra Group I Cytokine 27-plex (p 18) IL-2 Group l Cytokine 8-plex/ Group I Cytokine 8-plex/ Cytokine Th1/Th2 (p 37) 17-plex/27-plex/[Th1/Th2] (p 18) 23-plex/[Th1/Th2] (p 34) Chemokine 40-plex (p 21) IL-2Ra Group II Cytokine 21-plex (p 19) IL-3 Group II Cytokine 21-plex (p 19) Group I Cytokine 23-plex (p 34) IL-4 Group l Cytokine Group I Cytokine Cytokine Th1/Th2 (p 37) 8-plex/17-plex/27-plex/ 8-plex/23-plex/[Th1/Th2] (p 34) [Th1/Th2] (p 18) Chemokine 40-plex (p 21) IL-5 Group l Cytokine Group I Cytokine Cytokine Th1/Th2 (p 37) 17-plex/27-plex/[Th1/Th2] (p 18) 8-plex/23-plex/[Th1/Th2] (p 34) IL-6 Cancer 2 18-plex (p 25) Group I Cytokine Cytokine 24-plex/[Th1/Th2] (p 37) Group l Cytokine 8-plex/ 23-plex/[Th1/Th2]/Th17 (p 34) 17-plex/27-plex (p 18) Chemokine 40-plex (p 21) sil-6ra Cancer 1 16-plex (p 24) IL-7 Group l Cytokine 17-plex/27-plex (p 18) IL-8 / CXCL8 Cancer 2 18-plex (p 25) Group l Cytokine 8-plex/ 17-plex/27-plex (p 18) Chemokine 40-plex (p 21) IL-9 Group I Cytokine 27-plex (p 18) Group I Cytokine 23-plex (p 34) IL-10 Group l Cytokine 8-plex/ Group I Cytokine Cytokine 24-plex/[Th1/Th2] (p 37) 17-plex/27-plex/[Th1/Th2] (p 18) 8-plex/23-plex/ Chemokine 40-plex (p 21) [Th1/Th2]/ Th17 (p 34) IL-12 (p40) Group II Cytokine 21-plex (p 19) Group I Cytokine 23-plex (p 34) IL-12 (p70) Group l Cytokine 17-plex/ Group I Cytokine Cytokine 24-plex/[Th1/Th2] (p 37) 27-plex/[Th1/Th2] (p 18) 23-plex/[Th1/Th2] (p 34) IL-13 Group l Cytokine 17-plex/ Group I Cytokine 23-plex (p 34) Cytokine 24-plex/[Th1/Th2] (p 37) 27-plex/[Th1/Th2] (p 18) continues Bulletin

12 . Target Human (p 18 33) Mouse (p 34 36) Rat (p 37 39) Non-Human Primate (p 27) Canine (p 40) IL-15 Group I Cytokine 27-plex (p 18) Group II Cytokine 9-plex (p 35) IL-16 Group II Cytokine 21-plex (p 19) Chemokine 40-plex (p 21) IL-17A Group I Cytokine Cytokine 24-plex (p 37) 17-plex/27-plex (p 18) Th17 Cytokine 15-plex (p 20) IL-17A/F Th17 Cytokine singleplex (p 20) IL-17E / IL-25 Th17 Cytokine 15-plex (p 20) IL-17F Th17 Cytokine 15-plex (p 20) Group III Cytokine Th17 8-plex (p 35) IL-18 Cancer 2 18-plex (p 25) Group II Cytokine 9-plex (p 35) Kidney Toxicity 1 5-plex (p 39) Group II Cytokine 21-plex (p 19) Kidney Toxicity 1 6-plex (p 31) IL-21 Th17 Cytokine 15-plex (p 20) IL-22 Th17 Cytokine 15-plex (p 20) IL-23 Th17 Cytokine 15-plex (p 20) IL-27 (p28) Group III Cytokine (p 35) IL-33 Th17 Cytokine 15-plex (p 20) Insulin Diabetes 10-plex (p 27) Diabetes 8-plex (p 36) Diabetes 10-plex (p 27) IP-10 / CXCL10 Group I Cytokine 27-plex (p 18) Chemokine 40-plex (p 21) I-TAC / CXCL11 Chemokine 40-plex (p 21) KC Group I Cytokine 23-plex (p 34) KIM-1 Kidney Toxicity 1 6-plex (p 31) Kidney Toxicity 1 5-plex (p 39) Kidney Toxicity 1 4-plex (p 40) Lamin B, intact and 45 kd Apoptosis 1 (p 26) Leptin Cancer 1 16-plex (p 24) Diabetes 8-plex (p 36) Diabetes (p 38) Diabetes 10-plex (p 27) Diabetes 10-plex (p 27) sleptin R Metabolic Panel 2 (p 28) LH Hormone Panel 1 (p 28) LIF Group II Cytokine 21-plex (p 19) Group II Cytokine 9-plex (p 35) a-2-macroglobin Acute Phase 4-plex (p 33) Mcl-1 Apoptosis 2 (p 26) Mcl-1 / Bak dimer Apoptosis 3 (p 26) continues 2014 Bio-Rad Laboratories, Inc. 10

13 . Target Human (p 18 33) Mouse (p 34 36) Rat (p 37 39) Non-Human Primate (p 27) Canine (p 40) MCP-1 / CCL2 Group I Cytokine Group I Cytokine 23-plex (p 34) Kidney Toxicity 1 5-plex (p 39) Kidney Toxicity 1 4-plex (p 40) 17-plex/27-plex (p 18) Kidney Toxicity 1 6-plex (p 31) Chemokine 40-plex (p 21) MCP-2 / CCL8 Chemokine 40-plex (p 21) MCP-3 / CCL7 Group II Cytokine 21-plex (p 19) Chemokine 40-plex (p 21) MCP-4 / CCL13 Chemokine 40-plex (p 21) M-CSF Group II Cytokine 21-plex (p 19) Group II Cytokine 9-plex (p 35) MDC / CCL22 Chemokine 40-plex (p 21) MIF Group II Cytokine 21-plex (p 19) Chemokine 40-plex (p 21) MIG / CXCL9 Group II Cytokine 21-plex (p 19) Group II Cytokine 9-plex (p 35) Chemokine 40-plex (p 21) MIP-1a / CCL3 Group I Cytokine 27-plex (p 18) Group I Cytokine 23-plex (p 34) Chemokine 40-plex (p 21) MIP-1d / CCL15 Chemokine 40-plex (p 21) MIP-1b Group I Cytokine Group I Cytokine 23-plex (p 34) 17-plex/27-plex (p 18) MIP-2 Group II Cytokine 9-plex (p 35) Cytokine (p 37) MIP-3a / CCL20 Chemokine 40-plex (p 21) Group III Cytokine Cytokine 24-plex (p 37) Th17 8-plex (p 35) MIP-3b / CCL19 Chemokine 40-plex (p 21) MMP-1 MMPs and TIMPS (p 30) MMP-2 MMPs and TIMPS (p 30) MMP-3 MMPs and TIMPS (p 30) MMP-7 MMPs and TIMPS (p 30) MMP-8 MMPs and TIMPS (p 30) MMP-9 MMPs and TIMPS (p 30) MMP-10 MMPs and TIMPS (p 30) MMP-12 MMPs and TIMPS (p 30) MMP-13 MMPs and TIMPS (p 30) MPIF-1 / CCL23 Chemokine 40-plex (p 21) NGAL Kidney Toxicity 2 6-plex (p 31) Kidney Toxicity 2 4-plex (p 39) Kidney Toxicity 1 4-plex (p 40) b-ngf Group II Cytokine 21-plex (p 19) continues Bulletin

14 . Target Human (p 18 33) Mouse (p 34 36) Rat (p 37 39) Non-Human Primate (p 27) Canine (p 40) Omentin-1 Metabolic Panel 2 (p 28) Osteopontin Cancer 1 16-plex (p 24) Kidney Toxicity 1 5-plex (p 39) Kidney Toxicity 2 6-plex (p 31) PAI-1 Cancer 2 18-plex (p 25) Diabetes 8-plex (p 36) Diabetes 10-plex (p 27) Diabetes 10-plex (p 27) Pancreatic polypeptide Metabolic Panel 1 (p 28) PDGF-AB / BB Cancer 1 16-plex (p 24) Group II Cytokine 9-plex (p 35) Group I Cytokine 27-plex (p 18) PECAM-1 Cancer 1 16-plex (p 24) Pentraxin-3 Metabolic Panel 2 (p 28) Peptide YY Metabolic Panel 1 (p 28) PLGF Cancer 2 18-plex (p 25) PON-1 Metabolic Panel 2 (p 28) Procalcitonin Acute Phase 5-plex (p 33) Proinsulin Metabolic Panel 1 (p 28) Prolactin Cancer 1 16-plex (p 24) Hormone Panel 1 (p 28) Protein S Metabolic Panel 4 (p 28) RANTES Group I Cytokine 27-plex (p 18) Group I Cytokine 23-plex (p 34) RBP-4 Metabolic Panel 3 (p 28) Resistin Diabetes 10-plex (p 27) Diabetes 8-plex (p 36) Diabetes 10-plex (p 27) SAA Acute Phase 5-plex (p 33) SAP Acute Phase 4-plex (p 33) SCF Cancer 1 16-plex (p 24) Group II Cytokine 21-plex (p 19) SCGF-b Group II Cytokine 21-plex (p 19) SCYB16 / CXCL16 Chemokine 40-plex (p 21) SDF-1a+b / Group II Cytokine 21-plex (p 19) CXCL12 Chemokine 40-plex (p 21) SEPP Metabolic Panel 4 (p 28) Smac Apoptosis 1 (p 26) Survivin Apoptosis 3 (p 26) TARC / CCL17 Chemokine 40-plex (p 21) TECK / CCL25 Chemokine 40-plex (p 21) continues 2014 Bio-Rad Laboratories, Inc. 12

15 . Target Human (p 18 33) Mouse (p 34 36) Rat (p 37 39) Non-Human Primate (p 27) Canine (p 40) TFF3 Kidney Toxicity 2 6-plex (p 31) TGF-a Cancer 2 18-plex (p 25) TGF-b1 TGF-b (p 23) TGF-b (p 35) TGF-b (p 37) TGF-b2 TGF-b (p 23) TGF-b (p 35) TGF-b (p 37) TGF-b3 TGF-b (p 23) TGF-b (p 35) TGF-b (p 37) stie-2 Cancer 1 16-plex (p 24) TIMP-1 MMPs and TIMPS (p 30) TIMP-2 MMPs and TIMPS (p 30) TIMP-3 MMPs and TIMPS (p 30) TIMP-4 MMPs and TIMPS (p 30) TNF-a Cancer 2 18-plex (p 25) Group I Cytokine 8-plex/ Cytokine Th1/Th2 (p 37) Group I Cytokine 8-plex/ 23-plex/[Th1/Th17] (p 34) 17-plex/27-plex/[Th1/Th2] (p 18) Chemokine 40-plex (p 21) TNF-b Group II Cytokine 21-plex (p 19) tpa Acute Phase 5-plex (p 33) TRAIL Group II Cytokine 21-plex (p 19) Transferrin Metabolic Panel 3 (p 28) Transthyretin Metabolic Panel 3 (p 28) TSH Hormone Panel 1 (p 28) upa Cancer 2 18-plex (p 25) Vaspin Metabolic Panel 2 (p 28) VCAM-1 Group II Cytokine singleplex (p 19) VDBP Metabolic Panel 3 (p 28) VEGF Group I Cytokine singleplex (p 18) Group II Cytokine 9-plex (p 35) VEGF-A Cancer 2 18-plex (p 25) VEGF-C Cancer 2 18-plex (p 25) VEGF-D Cancer 2 18-plex (p 25) svegfr-1 Cancer 1 16-plex (p 24) svegfr-2 Cancer 1 16-plex (p 24) Visfatin Diabetes 10-plex (p 27) Diabetes 10-plex (p 27) Bulletin

16 Bio-Plex Magnetic Cell Signaling Assays Bio-Plex Pro Cell Signaling Phosphoprotein Assays bio-rad.com/cellsignalingassays Bio-Rad bulletin 6285 Lysate Controls Lysate Controls for Total Protein for Total Protein Phosphoprotein Singleplex Cat. Phosphoprotein Assays Lysate Cat. Singlplex Cat. Assays Lysate Cat. Bead Region Akt (Ser 473 ) 171-V50001M EGF-treated HEK YZ V60001M Untreated HeLa 171-YZT Akt (Thr 308 ) 171-V50002M EGF-treated HEK YZ V60001M Untreated HeLa 171-YZT ATF-2 (Thr 71 ) 171-V50024M UV-treated HEK YZ BAD (Ser 136 ) 171-V50025M PDGF-treated NIH3T3 171-YZ Btk (Tyr 223 ) 171-V50026M H 2 O 2 -treated Ramos 171-YZ V60012M H 2 O 2 -treated Ramos 171-YZ c-abi (Tyr 245 ) 171-V50027M Untreated K YZT c-jun (Ser 63 ) 171-V50003M UV-treated HEK YZ V60002M Untreated HEK YZT CREB (Ser 133 ) 171-V50028M UV-treated HEK YZ V60013M Untreated HEK YZT EGFR (Tyr 1068 ) 171-V50004M EGF-treated HeLa 171-YZ EGFR (Tyr 1173 ) 171-V50005M EGF-treated HeLa 171-YZ Erk1/2 (Thr 202 /Tyr 204, Thr 185 /Tyr 187 ) 171-V50006M EGF-treated HEK YZ V60003M Untreated HeLa 171-YZT GSK-3a/b (Ser 21 /Ser 9 ) 171-V50007M EGF-treated HEK YZ V60004M Untreated HeLa 171-YZT HER-2 (Tyr 1248 ) 171-V50008M EGF-treated SK-Br3 171-YZ V60005M EGF-treated SK-Br3 171-YZ HSP27 (Ser 78 ) 171-V50029M EGF-treated SK-Br3 171-YZ IGF-1R (Tyr 1131 ) 171-V50009M IGF-1-treated HEK YZ V60014M Untreated HEK YZT IR-b (Tyr 1146 ) 171-V50031M TNF-a-treated HeLa 171-YZ IRS-1 (Ser 636 /Ser 639 ) 171-V50030M PDGF-treated NIH3T3 171-YZ IkB-a (Ser 32 /Ser 36 ) 171-V50010M TNF-a-treated HeLa 171-YZ V0006M Untreated HeLa 171-YZT JNK (Thr 183 /Tyr 185 ) 171-V50011M UV-treated HEK YZ V0007M Untreated HeLa 171-YZT Lyn (Tyr 507 ) 171-V50032M H 2 O 2 -treated Ramos 171-YZ MEK1 (Ser 217 /Ser 221 ) 171-V50012M EGF-treated HEK YZ V0008M Untreated HeLa 171-YZT mtor (Ser 2448 ) 171-V50033M PDGF-treated NIH3T3 171-YZ V60015M Untreated HeLa 171-YZT NF-kB p65 (Ser 536 ) 171-V50013M TNF-a-treated HeLa 171-YZ p38 MAPK (Thr 180 /Tyr 182 ) 171-V50014M UV-treated HEK YZ V60009M Untreated HeLa 171-YZT p53 (Ser 15 ) 171-V50034M UV-treated HEK YZ p70 S6 Kinase (Thr 389 ) 171-V50015M NGFb-treated PC YZ V60010M Untreated HeLa 171-YZT p70 S6 Kinase (Thr 421 /Ser 424 ) 171-V50016M NGFb-treated PC YZ V60010M Untreated HeLa 171-YZT p90 RSK (Ser 380 ) 171-V50035M EGF-treated SK-Br3 171-YZ continues 2014 Bio-Rad Laboratories, Inc. 14

17 Bio-Plex Magnetic Cell Signaling Assays Bio-Plex Pro Cell Signaling Phosphoprotein Assays bio-rad.com/cellsignalingassays Bio-Rad bulletin 6285 Lysate Controls Lysate Controls Total Protein for Total Protein Phosphoprotein for Singleplex Cat. Phosphoprotein Assays Lysate Cat. Singlplex Cat. Assays Lysate Cat. Bead Region PDGFR-a (Tyr 754 ) 171-V50017M PDGF-treated NIH3T3 171-YZ PDGFR-b (Tyr 751 ) 171-V50018M PDGF-treated NIH3T3 171-YZ Pl3K p85 (Tyr 458 ) 171-V50036M H 2 O 2 -treated Ramos 171-YZ PTEN (Ser 380 ) 171-V50037M PDGF-treated NIH3T3 171-YZ V60016M Untreated HeLa 171-YZT S6 ribosomal protein (Ser 235 /Ser 236 ) 171-V50038M EGF-treated SK-Br3 171-YZ Smad2 (Ser 465 /Ser 467 ) 171-V50019M TNF-a-treated HeLa 171-YZ V60011M Untreated HeLa 171-YZT Src (Tyr 416 ) 171-V50039M Src-transfected NIH3T3 171-YZ V60017M Src-transfected NIH3T3 171-YZ Stat1 (Tyr 701 ) 171-V50020M IFN-a-treated HeLa 171-YZ Stat3 (Ser 727 ) 171-V50021M IFN-a-treated HeLa 171-YZ Stat3 (Tyr 705 ) 171-V50022M IFN-a-treated HeLa 171-YZ Syk (Tyr 352 ) 171-V50040M H 2 O 2 -treated Ramos 171-YZ VEGFR-2 (Tyr 1175 ) 171-V50023M VEGF-treated HUVEC 171-YZ ZAP-70 (Tyr 319 ) 171-V50041M H 2 O 2 -treated Jurkat 171-YZ V60018M H 2 O 2 -treated Jurkat 171-YZ House Keeping Protein Singleplex Cat. Lysate Control Catalog Human GAPDH 171-V60019M Untreated HeLa 171-YZT002 b-actin 171-V60020M Untreated HeLa 171-YZT002 Negative control for all negative control for all house keeping Detection antibody diluent* protein assays Negative Controls Lysate Control Catalog Negative control for all phosphoprotein assays Phosphatase-treated HeLa 171-YZB001 Negative control for all total target assays Detection antibody diluent* * Detection antibody diluent is included with the Bio-Plex Pro Cell Signaling Reagent Kit (cat # M). Bio-Plex Pro Cell Signaling Akt Panel 8-Plex (LQ-00006JK0K0RR) Analyte Akt (Ser 473 ) mtor (Ser 2448 ) BAD (Ser 136 ) p70 S6 Kinase (Thr 389 ) GSK-3a/b (Ser 21 /Ser 9 ) PTEN (Ser 380 ) IRS-1 (Ser 636 /Ser 639 ) S6 ribosomal protein (Ser 235 /Ser 236 ) Bio-Plex Pro Cell Signaling MAPK Panel 9-plex (LQ-00000S6KL81S) Analyte ATF2 (Thr 71 ) p38 (Thr 180 /Tyr 182 ) ERK1/2 (Thr 202 /Tyr 204 Thr 185 /Tyr 187 ) p53 (Ser 15 ) HSP27 (Ser 78 ) p90 RSK (Ser 380 ) JNK (Thr 183 /Tyr 185 ) Stat3 (Ser 727 ) MEK1 (Ser 217 /Ser 221 ) Bulletin

18 Bio-Plex Magnetic Cell Signaling Assays Phosphoprotein Assays P/B Ratio* Intra-Assay %CV Inter-Assay %CV Sensitivity (LOD), µg/well Reaction Species** Akt (Ser 473 ) H, M, R Akt (Thr 308 ) H, M, R ATF-2 (Thr 71 ) H, M, R, NHP BAD (Ser 136 ) H, M, R, NHP Btk (Tyr 223 ) H c-abi (Tyr 245 ) H, M c-jun (Ser 63 ) H, M, R CREB (Ser 133 ) H, M, R EGFR (Tyr 1068 ) H EGFR (Tyr 1173 ) H Erk1/2 (Thr 202 /Tyr 204, Thr 185 /Tyr 187 ) H, M GSK-3a/b (Ser 21 /Ser 9 ) H, M, R HER-2 (Tyr 1248 ) H, M, R HSP27 (Ser 78 ) H, NHP IGF-1R (Tyr 1131 ) H IR-b (Tyr 1146 ) H, M, R IRS-1 (Ser 636 /Ser 639 ) H, M, R IkB-a (Ser 32 /Ser 36 ) H JNK (Thr 183 /Tyr 185 ) H Lyn (Tyr 507 ) H, M, R MEK1 (Ser 217 /Ser 221 ) H, M, R mtor (Ser 2448 ) H, M, R, NHP NF-kB p65 (Ser 536 ) H, M, R p38 MAPK (Thr 180 /Tyr 182 ) H, M, R p53 (Ser 15 ) H p70 S6 Kinase (Thr 389 ) H, M, R p70 S6 Kinase (Thr 421 /Ser 424 ) H, M, R p90 RSK (Ser 380 ) H, M, R, NHP PDGFR-a (Tyr 754 ) H, M PDGFR-b (Tyr 751 ) H, M, R Pl3K p85 (Tyr 458 ) H, M, R PTEN (Ser 380 ) H, M, R continues Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bead Type 6.5 µm Magnetic bio-rad.com/cellsignalingassays Bio-Rad bulletin Bio-Rad Laboratories, Inc. 16

19 Bio-Plex Magnetic Cell Signaling Assays bio-rad.com/cellsignalingassays Bio-Rad bulletin 6285 Phosphoprotein Assays P/B Ratio* Intra-Assay %CV Inter-Assay %CV Sensitivity (LOD), µg/well Reaction Species** S6 Ribosomal Protein (Ser 235 /Ser 236 ) H, M, R Smad2 (Ser 465 /Ser 467 ) H, M, R Src (Tyr 416 ) H, M, R Stat1 (Tyr 701 ) H Stat3 (Ser 727 ) H, M, R Stat3 (Tyr 705 ) H, M, R Syk (Tyr 352 ) H, M, R VEGFR-2 (Tyr 1175 ) H, M ZAP-70 (Tyr 319 ) H, M * Signal ratio (positive/background) of the stimulated cell lysate divided by the negative control cell lysate. ** H, human; M, mouse; R, rat; NHP, non-human primate Total Target Assays Intra-Assay %CV Inter-Assay %CV Sensitivity (LOD), µg/well Reaction Species* b-actin H, M, R, NHP Akt H, M, R Btk H, M c-jun H CREB H, M, R, NHP Erk1/ H, M GAPDH H GSK-3b H, M, R HER H, M, R IGF-1R H IkB-a H JNK H MEK H, M, R mtor H, M, R, NHP p38 MAPK H, M, R p70 S6 Kinase H, M, R PTEN H, M, R Smad H Src H, M, R ZAP H * H, human; M, mouse; R, rat; NHP, non-human primate. Bulletin

20 Human Assays Bio-Plex Pro Human Cytokine Group I Assays bio-rad.com/ccgfassays Bio-Rad bulletin 5828 Bio-Plex Pro Human Group l Analytes Eotaxin 171-B5015M 43 FGF-basic 171-B5016M 44 G-CSF 171-B5017M 57 GM-CSF 171-B5018M 34 IFN-g 171-B5019M 21 IL-b 171-B5001M 39 IL-1Ra 171-B5002M 25 IL B5003M 38 IL B5004M 52 IL B5005M 33 IL B5006M 19 IL B5007M 74 IL B5008M 54 IL B5009M 77 IL B5010M 56 IL-12 (p70) 171-B5011M 75 IL B5012M 51 IL B5013M 73 IL-17A 171-B5014M 76 IP B5020M 48 MCP-1 (MCAF) 171-B5021M 53 MIP-1a 171-B5022M 55 MIP-1b 171-B5023M 18 PDGF-BB 171-B5024M 47 RANTES 171-B5025M 37 TNF-a 171-B5026M 36 VEGF 171-B5027M 45 Bio-Plex Pro Human Cytokine 8-Plex Panel (M A) GM-CSF IL-6 IFN-g IL-8 IL-2 IL-10 IL-4 TNF-a Bio-Plex Pro Human Cytokine Th1/Th2 Panel (M L3) GM-CSF IL-10 IFN-g IL-12 (p70) IL-2 IL-13 IL-4 TNF-a IL-5 Bio-Plex Pro Human Cytokine 17-Plex Panel (M YV) G-CSF IL-8 GM-CSF IL-10 IFN-g IL-12 (p70) IL-1b IL-13 IL-2 IL-17A IL-4 MCP-1 (MCAF) IL-5 MIP-1b IL-6 TNF-a IL-7 Bio-Plex Pro Human Cytokine 27-Plex Panel (M50-0KCAF0Y) Eotaxin IL-9 FGF-basic IL-10 G-CSF IL-12p70 GM-CSF IL-13 IFN-g IL-15 IL-1b IL-17A IL-1Ra IP-10 IL-2 MCP-1 (MCAF) IL-4 MIP-1a IL-5 MIP-1b IL-6 PDGF-BB IL-7 RANTES IL-8 TNF-a Compatible with These Readers Bead Type Bio-Plex MAGPIX Luminex MAGPIX 6.5 µm Bio-Plex 200 Luminex 200 Magnetic Bio-Plex 3D Luminex FLEXMAP 3D Assay Working Range, Assay Sensitivity, pg/ml pg/ml Assay Precision Target LLOQ ULOQ LOD Intra-Assay %CV Inter-Assay %CV Group I 27-Plex Panel Eotaxin , FGF-basic , G-CSF , GM-CSF , IFN-g , IL-1b 3.2 3, IL-1Ra , IL , IL , IL , IL , IL , IL , IL , IL , IL-12 (p70) , IL , IL , IL-17A , IP , MCP-1 (MCAF) 2.1 1, MIP-1a MIP-1b 2.0 1, PDGF-BB , RANTES 2.2 8, TNF-a , VEGF , Bio-Rad Laboratories, Inc. 18

21 Bio-Plex Pro Human Cytokine Group II Assays bio-rad.com/ccgfassays Bio-Rad bulletin 5828 Bio-Plex Pro Human Cytokine Group ll Analytes CTACK 171-B6006M 72 Gro-a 171-B6007M 61 HGF 171-B6008M 62 ICAM-1** 171-B6009M 12 IFN-a2 171-B6010M 20 IL-1a 171-B6001M 63 IL-2Ra 171-B6002M 13 IL B6003M 64 IL-12 (p40) 171-B6004M 28 IL B6005M 27 IL-18* 42 LIF 171-B6011M 29 MCP B6012M 26 M-CSF 171-B6013M 67 MIF 171-B6014M 35 MIG 171-B6015M 14 b-ngf 171-B6016M 46 SCF 171-B6017M 65 SCGF-b 171-B6018M 78 SDF-1a+b 171-B6019M 22 TNF-b 171-B6020M 30 TRAIL 171-B6021M 66 VCAM-1** 171-B6022M 15 * Not available as a singleplex. ** ICAM-1 and VCAM-1 can be multiplexed together but not with other analytes. Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bio-Plex Pro Human Cytokine Group ll 21-Plex Panel (MF0-005KMII) CTACK MCP-3 Gro-a M-CSF HGF MIF IFN-a2 MIG IL-1a b-ngf IL-2Ra SCF IL-3 SCGF-b IL-12 (p40) SDF-1a+b IL-16 TNF-b IL-18 TRAIL LIF Bead Type 6.5 µm Magnetic Assay Working Range, Assay Sensitivity, pg/ml pg/ml Assay Precision Target LLOQ ULOQ LOD Intra-Assay %CV Inter-Assay %CV Group II 21-Plex Panel, ICAM-1 and VCAM-1 CTACK 13 16, Gro-a 19 2, HGF , ICAM , IFN-a , IL-1a , IL-2Ra 299 7, IL , IL-12 (p40) 41 28, IL , IL , LIF 12 22, MCP , M-CSF , MIF , MIG 2.7 8, b-ngf 1.0 4, SCF , SCGF-b 66 47, SDF-1a+b 10 10, TNF-b , TRAIL , VCAM , Bulletin

22 Bio-Plex Pro Human Th17 Cytokine bio-rad.com/ccgfassays Bio-Rad bulletin 6249 Bio-Plex Pro Human Th17 Analytes IFN-g 171-BA013M 21 IL-1b 171-BA001M 39 ll BA002M 52 ll BA003M 19 IL BA004M 56 IL-17A 171-BA005M 76 IL-17A /F 171-BA016M 66 IL-17F 171-BA006M 44 IL BA007M 47 IL BA008M 18 IL BA009M 43 IL BA010M 55 IL BA011M 62 IL BA012M 46 scd40l 171-BA014M 29 TNF-a 171-BA015M 36 Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bio-Plex Pro Human Th17 Cytokine 15-Plex Panel (171-AA001M) IFN-g IL-21 IL-1b IL-22 ll-4 IL-23 ll-6 IL-25 IL-10 IL-31 IL-17A IL-33 IL-17F scd40l Bead Type 6.5 µm Magnetic Human Th17 Analytes Assay Working Assay Sensitivity, Assay Calibration to Range, pg/ml pg/ml Precision WHO/NIBSC Alternate Intra-Assay Inter-Assay NIBSC Target Names LLOQ ULOQ LOD %CV %CV Factor** Catalog # IFN-g , /586 IL-1b , /680 IL , /656 IL , /548 IL , /722 IL-17A IL-17, CTLA , /420 IL-17A/F* , IL-17F , IL-21 Za , IL , IL-23 IL-23A, SGRF , IL-25 IL-17E , IL , IL , scd-40l CD , TNF-a , /786 * IL-17A/F values are derived from the singleplex assay format. ** Factor NIBSC value (pg/ml) = Bio-Plex value (pg/ml). Factors are based on average % recovery of Bio-Plex standards (in the linear range) relative to NIBSC standards run on the same assay plate Bio-Rad Laboratories, Inc. 20

23 Bio-Plex Pro Human Chemokine Assays bio-rad.com/chemokineassays Bio-Rad bulletin 6499 Bio-Plex Pro Human Chemokine Panel, 40-Plex (171-AK99MR2) 6Ckine / CCL BK11MR2 12 BCA-1 / CXCL BK12MR2 74 CTACK / CCL BK13MR2 72 ENA-78 / CXCL5 171-BK14MR2 73 Eotaxin / CCL BK15MR2 43 Eotaxin-2 / CCL BK16MR2 30 Eotaxin-3 / CCL BK17MR2 65 Fractalkine / CX3CL1 171-BK18MR2 77 GCP-2 / CXCL6 171-BK19MR2 15 GM-CSF 171-BK21MR2 34 Gro-a / CXCL1 171-BK22MR2 61 Gro-b / CXCL2 171-BK23MR2 78 I-309 / CCL1 171-BK24MR2 20 IFN-g 171-BK25MR2 21 IL-1b 171-BK26MR2 39 IL BK27MR2 38 IL BK28MR2 52 IL BK29MR2 19 IL-8 / CXCL8 171-BK31MR2 54 IL BK32MR2 56 IL BK33MR2 27 IP-10 / CXCL BK34MR2 48 I-TAC / CXCL BK35MR2 25 MCP-1 / CCL2 171-BK36MR2 53 MCP-2 / CCL8 171-BK37MR2 57 MCP-3 / CCL7 171-BK38MR2 26 MCP-4 / CCL BK39MR2 28 MDC / CCL BK41MR2 29 MIF 171-BK42MR2 35 MIG / CXCL9 171-BK43MR2 14 MIP-1a / CCL3 171-BK44MR2 55 MIP-1d / CCL BK46MR2 66 MIP-3a / CCL BK47MR2 62 MIP-3b / CCL BK48MR2 76 MPIF-1 / CCL BK49MR2 37 SCYB16 / CXCL BK51MR2 64 SDF-1a+b / CXCL BK52MR2 22 TARC / CCL BK53MR2 67 TECK / CCL BK54MR2 46 TNF-a 171-BK55MR2 36 Compatible with These Readers Bead Type Bio-Plex MAGPIX Luminex MAGPIX 6.5 µm Bio-Plex 200 Luminex 200 Magnetic Bio-Plex 3D Luminex FLEXMAP 3D Human Chemokines Assay Working Ranges, pg/ml Assay Sensitivity Assay Precision Calibration to WHO/NIBSC Analyte LLOQ ULOQ LOD Intra-Assay %CV Inter-Assay %CV Factor* Catalog # 6Ckine / CCL , BCA-1 / CXCL , CTACK / CCL , ENA-78 / CXCL , Eotaxin / CCL , Eotaxin-2 / CCL , Eotaxin-3 / CCL , Fractalkine / CX3CL , continues Bulletin

24 Bio-Plex Pro Human Chemokine Assays bio-rad.com/chemokineassays Bio-Rad bulletin 6499 Human Chemokines Assay Working Ranges, pg/ml Assay Sensitivity Assay Precision Calibration to WHO/NIBSC Analyte LLOQ ULOQ LOD Intra-Assay %CV Inter-Assay %CV Factor* Catalog # GCP-2 / CXCL , GM-CSF , Gro-a / CXCL , Gro-b / CXCL , I-309 / CCL , IFN-g , /586 IL-1b 0.4 7, /680 IL , IL , /656 IL , /548 IL-8 / CXCL , /520 IL , /722 IL , IP-10 / CXCL , I-TAC / CXCL , MCP-1 / CCL , /794 MCP-2 / CCL , MCP-3 / CCL , MCP-4 / CCL , MDC / CCL , MIF , MIG / CXCL , MIP-1a / CCL , /518 MIP-1d / CCL , MIP-3a / CCL , MIP-3b / CCL , MPIF-1 / CCL , SCYB16 / CXCL , SDF-1a+b / CXCL , TARC / CCL TECK / CCL , TNF-a , /786 * Factor x NIBSC value (pg/ml) = Bio-Plex value (pg/ml). Factors are based on average % recovery of the Bio-Plex standards (in the linear range) relative to NIBSC standards run on the same assay plate Bio-Rad Laboratories, Inc. 22

25 TGF-b Assays Bio-Plex Pro TGF-b 3-Plex Panel (171-W4001M) TGF-b1 171-V4001M 13 TGF-b2 171-V4002M 72 TGF-b3 171-V4003M 66 Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bead Type 6.5 µm Magnetic Human TGF-b 3-Plex Panel Assay Working Range, Assay Sensitivity, pg/ml pg/ml Assay Precision Target LLOQ ULOQ LOD Intra-Assay %CV Inter-Assay %CV Premixed 3-Plex Assay bio-rad.com/ccgfassays Bio-Rad bulletin 6054 TGF-b , TGF-b , TGF-b , Bulletin

26 Bio-Plex Pro Human Cancer Biomarker Panels bio-rad.com/bio-plexprocancer Bio-Rad bulletin 6159 Bio-Plex Pro Human Cancer Biomarker Panel 1 16-plex (171-AC500M) Receptors, cytokines, chemokines, growth factors, and hormones segfr 171-BC501M 15 FGF-basic 171-BC502M 44 Follistatin 171-BC503M 26 G-CSF 171-BC504M 57 HGF 171-BC505M 62 sher-2/neu 171-BC506M 12 sil-6ra 171-BC507M 19 Leptin 171-BC508M 78 Osteopontin 171-BC509M 77 PECAM BC510M 47 PDGF-AB/BB 171-BC511M 46 Prolactin 171-BC512M 52 SCF 171-BC513M 65 stie BC514M 64 svegfr BC515M 76 svegfr BC516M 45 Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bead Type 6.5 µm Magnetic Human Cancer Biomarker Panel 1 Receptors, cytokines, chemokines, growth factors, and hormones Assay Working Assay Sensitivity, Assay Calibration to Range, pg/ml pg/ml Precision WHO/NIBSC Alternate Intra-Assay Inter-Assay NIBSC Target Names LLOQ ULOQ LOD %CV %CV Factor* Catalog # segfr ErbB1, EGFR , FGF-basic FGF2, BFGF , /712 Follistatin FSN , G-CSF , /136 HGF SF , /564 sher-2/neu ErbB , sil-6ra CD , Leptin , /594 Osteopontin OPN , PECAM-1 CD , PDGF-AB/BB , /728 Prolactin PRL , /582 SCF KL , /682 stie-2 CD202b , svegfr-1 FLT , svegfr-2 KDR, FLK , * Factor NIBSC value (pg/ml) = Bio-Plex value (pg/ml). Factors are based on average % recovery of Bio-Plex standards (in the linear range) relative to NIBSC standards run on the same assay plate Bio-Rad Laboratories, Inc. 24

27 Bio-Plex Pro Human Cancer Biomarker Panels bio-rad.com/bio-plexprocancer Bio-Rad bulletin 6159 Bio-Plex Pro Human Cancer Biomarker Panel 2 18-plex (171-AC600M) Ligands for receptors in panel 1 cytokines, chemokines, and growth factors Angiopoietin BC601M 47 scd40l 171-BC602M 29 EGF 171-BC603M 14 Endoglin 171-BC604M 26 sfasl 171-BC605M 35 HB-EGF 171-BC606M 57 IGFBP BC607M 62 IL BC608M 19 IL BC609M 54 IL-18* 42 PAI BC611M 61 PLGF 171-BC612M 52 TGF-a 171-BC613M 76 TNF-a 171-BC614M 36 upa 171-BC615M 78 VEGF-A 171-BC616M 45 VEGF-C 171-BC617M 64 VEGF-D 171-BC618M 43 * IL-18 is not available in singleplex format Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bead Type 6.5 µm Magnetic Human Cancer Biomarker Panel 2 Ligands for receptors in panel 1 cytokines, chemokines, and growth factors Assay Working Assay Sensitivity, Assay Calibration to Range, pg/ml pg/ml Precision WHO/NIBSC Alternate Intra-Assay Inter-Assay NIBSC Target Names LLOQ ULOQ LOD %CV %CV Factor* Catalog # Angiopoietin , scd40l TRAP , EGF 0.3 4, /530 Endoglin CD , sfasl CD95L/APO-1L , HB-EGF 0.3 4, IGFBP , IL , /548 IL-8 CXCL , /520 IL , /200 PAI-1 SERPINE , PLGF PGF 0.6 9, /272 TGF-a 0.6 7, TNF-a 0.6 9, /786 upa Urokinase , VEGF-A VPF , /286 VEGF-C Flt4 ligand , VEGF-D FIGF , * Factor NIBSC value (pg/ml) = Bio-Plex value (pg/ml). Factors are based on average % recovery of Bio-Plex standards (in the linear range) relative to NIBSC standards run on the same assay plate. Bulletin

28 Bio-Plex RBM Apoptosis Assays Bio-Plex Pro RBM Apoptosis Panel 1 (171-WAR1CK) Analyte Bead Region Bak 74 Bax 27 Lamin B, intact and 45 kd 14 Smac 19 Bio-Plex Pro RBM Apoptosis Panel 2 (171-WAR2CK) Analyte Bead Region Bad 73 Bax / Bcl-2 dimer 42 Bcl-xL 22 Bim 12 Mcl-1 18 Bio-Plex Pro RBM Apoptosis Panel 3 (171-WAR3CK) Analyte Bead Region Bcl-xL / Bak dimer 47 Caspase-3, active 57 Mcl-1 / Bak dimer 54 Survivin 20 bio-rad.com/apoptosisassays Bio-Rad bulletin 6474 Apoptosis Assay Working Range, ng/ml Assay Sensitivity, ng/ml Assay Precision Target Bead Region LLOQ ULOQ LOD Intra-Assay %CV Inter-Assay %CV Panel 1 Bak % 14% Bax % 19% Lamin B, intact and 45 kd % 14% Smac % 19% Panel 2 Bad % 9% Bax/Bcl-2 dimer % 9% Bcl-xL % 7% Bim % 8% Mcl % 6% Panel 3 Bcl-xL/Bak dimer % 14% Caspase-3, active % 8% Mcl-1/Bak dimer % 13% Survivin % 8% Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bead Type 6.5 µm Magnetic 2014 Bio-Rad Laboratories, Inc. 26

29 Bio-Plex Pro Human Diabetes Assays (compatible with Non-Human Primate samples) bio-rad.com/bio-plexprodiabetes Bio-Rad bulletin 6342 Bio-Plex Pro Human Diabetes 10-Plex Panel (171-A7001M) C-Peptide 171-B7003M 72 Ghrelin 171-B7004M 26 GIP 171-B7005M 14 GLP B7006M 27 Glucagon 171-B7007M 15 Insulin 171-B7008M 12 Leptin 171-B7009M 78 PAI-1 (total) 171-B7010M 61 Resistin 171-B7011M 65 Visfatin 171-B7012M 22 Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bio-Plex Pro Human Diabetes Adipsin Assay All-in-One Kit (171-A7004M) Adipsin* 171-A7004M 35 Bio-Plex Pro Human Diabetes 2-Plex Panel (171-A7002M) Adiponectin* 171-A7003M 64 Adipsin* 171-A7004M 35 Bio-Plex Pro Human Diabetes Adiponectin Assay All-in-One Kit (171-A7003M) Adiponectin* 171-A7003M 64 * Adiponectin and adipsin can also be assayed; they are available as either singleplexes or a 2-plex. Due to different dilution schemes, these biomarkers should be assayed separately from the ten markers in the panel. Bead Type 6.5 µm Magnetic Human Diabetes Assay Working Range, Assay Sensitivity, pg/ml pg/ml Assay Precision Target LLOQ ULOQ LOD Intra-Assay%CV Inter-Assay%CV 2-Plex Assay Adiponectin , Adipsin 43 14, Plex Assay C-Peptide , Ghrelin , GIP , GLP , Glucagon , Insulin 1.7 3, Leptin , PAI , Resistin 2.3 4, Visfatin , Bulletin

30 Bio-Plex Pro Human Metabolic and Hormone Assays New bio-rad.com/bio-plexmetabolicassays Bio-Rad bulletin 6571 Bio-Plex Pro Human Metabolic Panel 1 (171-AMR1CK) Bio-Plex Pro Human Metabolic Panel 3 (171-AMR3CK) Bio-Plex Pro Human Hormone Panel 1 (171-AHR1CK) Bio-Plex Pro Human IGFBP Panel (171-AGR1CK) Analyte Bead Region Analyte Bead Region Analyte Bead Region Analyte Bead Region C-Peptide 45 Cortisol 12 Pancreatic polypeptide 52 Proinsulin 67 Peptide YY 55 Bio-Plex Pro Human Metabolic Panel 2 (171-AMR2CK) Analyte Bead Region FGF FGF Galectin-3 46 sleptin R 27 Omentin-1 20 Pentraxin-3 44 PON-1 53 Vaspin 34 AAT 26 AGP-1 57 Hemopexin 35 RBP-4 43 Transferrin 61 Transthyretin 13 VDBP 28 Bio-Plex Pro Human Metabolic Panel 4 (171-AMR4CK) Analyte Bead Region ANGPTL3 66 Chemerin 42 DPP4 25 Protein S 62 SEPP 36 FSH 18 GH 38 LH 15 Prolactin 19 TSH 22 Bio-Plex Pro Human IGF Panel (171-AFR1CK) Analyte Bead Region IGF-1 21 IGF-2 54 Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D IGFBP-1 48 IGFBP-2 65 IGFBP-3 33 IGFBP-4 76 IGFBP-5 51 IGFBP-6 78 IGFBP-7 37 Bead Type 6.5 µm Magnetic Assay Working Ranges Assay Sensitivity Assay Precision Panel Analyte Alternative Names Bead Region LLOQ ULOQ LOD Units Intra-Assay %CV Inter-Assay %CV Human C-Peptide Connecting peptide , pg/ml 2 5 Metabolic Cortisol 12 2, , pg/ml Panel 1 Pancreatic polypeptide PP pg/ml 3 5 Proinsulin , pg/ml 3 5 Peptide YY PYY , pg/ml 3 6 Human FGF-21 Fibroblast growth factor , pg/ml Metabolic FGF-23 Fibroblast growth factor , pg/ml Panel 2 Galectin-3 Galactoside-binding, soluble, , pg/ml sleptin R Soluble leptin receptor , pg/ml 8 9 Omentin-1 Intelectin ,277 13,500,000 6,407 pg/ml Pentraxin-3 PTX , pg/ml 7 12 PON-1 Serum paraoxonase ,343 12,150,000 6,300 pg/ml 8 8 Vaspin Visceral adipose tissue-derived serine protease inhibitor , pg/ml continues 2014 Bio-Rad Laboratories, Inc. 28

31 Assay Working Ranges Assay Sensitivity Assay Precision Panel Analyte Alternative Names Bead Region LLOQ ULOQ LOD Units Intra-Assay %CV Inter-Assay %CV Human AAT Alpha-1 antitrypsin ng/ml 6 7 Metabolic AGP-1 Alpha-1-acid glycoprotein ng/ml 7 11 Panel 3* Hemopexin ng/ml 8 8 RBP-4 Retinol-binding protein ng/ml 6 9 Transferrin Serotransferrin ng/ml 4 6 Transthyretin TTR ng/ml 6 6 VDBP Vitamin D-binding protein ng/ml 4 7 Human ANGPTL3 Angiopoietin-like ng/ml 8 10 Metabolic Chemerin ng/ml 5 9 Panel 4 DPP4 Dipeptidyl peptidase ng/ml 7 8 Protein S Vitamin K-dependent protein S ng/ml 8 9 SEPP Selenoprotein P , ng/ml 9 16 Human FSH Follicle-stimulating hormone miu/ml 7 8 Hormone GH Human growth hormone ng/ml 6 7 Panel 1 LH Luteinizing hormone miu/ml 4 6 Prolactin PRL ng/ml 7 8 TSH Thyroid-stimulating hormone µiu/ml 5 7 Human IGFBP-1 Insulin-like growth ng/ml 4 7 IGFBP IGFBP-2 factor binding proteins ng/ml 6 12 Panel IGFBP , ng/ml 4 13 IGFBP ng/ml 4 11 IGFBP ng/ml 5 14 IGFBP ng/ml 7 11 IGFBP ng/ml 5 9 Human IGF-1 Insulin-like growth factor ng/ml 2 6 IGF Panel IGF ng/ml 2 4 * The product datasheet supplied with metabolic panel 3 provides performance values as well as standard S1 and control values in μg/ml. Values here are in ng/ml for display purposes. Bulletin

32 Bio-Plex Pro Human MMP and TIMP Assays New bio-rad.com/bio-plexmmpsandtimps Bio-Rad bulletin 6551 Bio-Plex Pro Human MMP Panel 9-Plex (171-AM001M) MMP BM001M 43 MMP BM002M 26 MMP BM003M 45 MMP BM004M 66 MMP BM005M 37 MMP BM006M 55 MMP BM007M 62 MMP BM008M 53 MMP BM009M 47 Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bio-Plex Pro Human TIMP Panel 4-Plex (171-A002M) Analyte Bead Region TIMP-1 21 TIMP-2 64 TIMP-3 52 TIMP-4 35 Bead Type 6.5 µm Magnetic Bio-Plex Pro RBM Human MMP and TIMP Assays Assay Working Ranges, pg/ml Assay Sensitivity, pg/ml Analyte Bead Region LLOQ ULOQ LOD Human MMP Panel 9-Plex MMP , MMP ,357, MMP , MMP , MMP , MMP , MMP , MMP ,655 1 MMP , Human TIMP Panel 4-Plex TIMP , TIMP , TIMP , TIMP , Bio-Rad Laboratories, Inc. 30

33 Bio-Plex Pro Human Kidney Toxicity Assays bio-rad.com/bio-plexkidneytox Bio-Rad bulletin 6347 Bio-Plex Pro RBM Human Kidney Toxicity Panel 1 (171-ATR1CK) Analyte Bead Region Calbindin 64 Clusterin 12 GST-π 25 IL KIM-1 44 MCP-1 15 Compatible with These Readers Bio-Plex MAGPIX Luminex MAGPIX Bio-Plex 200 Luminex 200 Bio-Plex 3D Luminex FLEXMAP 3D Bio-Plex Pro RBM Human Kidney Toxicity Panel 2 (171-ATR2CK) Analyte Bead Region Albumin 30 B2M 22 Cystatin C 51 NGAL 46 Osteopontin 20 TFF3 61 Bead Type 6.5 µm Magnetic Kidney Toxicity Assay Working Assay Assay Precision Alternate Bead Ranges, ng/ml Sensitivity, ng/ml Intra-Assay Inter-Assay Analyte Names Region LLOQ ULOQ LOD %CV %CV Human Kidney Toxicity Panel 1 Calbindin , Clusterin ApoJ , GST-π IL KIM-1 TIM MCP Human Kidney Toxicity Panel 2 Albumin B2M Cystatin C NGAL Lipocalin Osteopontin OPN , TFF Bulletin

34 Bio-Plex Pro Human Isotyping Assays Bio-Plex Pro Human Isotyping Panel (171-A3100M) Analyte 6-Plex Singleplex Bead Region IgA 171-A3101M 12 IgM 171-A3104M 56 IgE* 171-A3102M 38 IgG-total* 171-A3103M 66 IgG 1 54 IgG 2 21 IgG 3 36 IgG 4 72 * IgE and IgG-total are not part of the 6-plex. Compatible with These Readers Bead Type Bio-Plex MAGPIX Luminex MAGPIX 6.5 µm Bio-Plex 200 Luminex 200 Magnetic Bio-Plex 3D Luminex FLEXMAP 3D bio-rad.com/bio-plexproisotyping Bio-Rad bulletin 6344 Human Isotyping Assay Working Range, Assay Sensitivity, pg/ml pg/ml Assay Precision Target LLOQ ULOQ LOD Intra-Assay%CV Inter-Assay%CV Singleplex Assay lge lgg total , Plex Assay lgg , lgg , lgg lgg lga lgm , Bio-Rad Laboratories, Inc. 32

35 Bio-Plex Pro Human Acute Phase Assays bio-rad.com/bio-plexproacutephase Bio-Rad bulletin 5650 Bio-Plex Pro Human Acute Phase Plex Panel (171-A4S07M) Analyte CRP Ferritin Fibrinogen Haptoglobin a-2-macroglobin Procalcitonin SAA SAP tpa Bio-Plex Pro Human Acute Phase 4-Plex Panel (171-A4C09M) Analyte CRP Haptoglobin a-2-macroglobin SAP Bio-Plex Pro Human Acute Phase 5-Plex Panel (171-A4C07M) Analyte Ferritin Fibrinogen Procalcitonin SAA tpa Human Acute Phase Assay Working Assay Target Range Sensitivity Dynamic WHO/NIBSC LLOQ ULOQQ LOD Ranges* Catalog # 1 IU = x ng** CRP 0.01 ng/ml 50 ng/ml ng/ml ng/ml 85/ x 10 6 Ferritin 3.05 pg/ml 50,000 pg/ml 1.3 pg/ml ,000 pg/ml Fibrinogen 5 ng/ml 813 ng/ml 2.8 ng/ml ,000 ng/ml Haptoglobin 0.1 ng/ml 500 ng/ml 0.07 ng/ml ng/ml a-2-macroglobulin 0.5 ng/ml 1,875 ng/ml 0.25 ng/ml 0.5 7,500 ng/ml Procalcitonin 14 pg/ml 10,000 pg/ml 11.3 pg/ml ,000 pg/ml SAA 1 ng/ml 700 ng/ml 1.1 ng/ml ,000 ng/ml Serum amyloid P 0.1 ng/ml 250 ng/ml ng/ml ng/ml (SAP) Tissue plasminogen 28 pg/ml 5,000 pg/ml 6.15 pg/ml ,000 pg/ml 98/ x 10 1 activator * Standard curve ranges. **IU, international units. Compatible with These Readers Bio-Plex 200 Luminex 200 Bead Type 8.0 µm Magnetic * Note: This panel is not compatible with the MAGPIX and 3D readers. Bulletin