Introduction
The Human Protein Phosphorylation Antibody Array 1 is specifically designed for simultaneous identification of the relative levels of phosphorylation of 507 different human proteins in lysate samples. By monitoring the changes in protein phosphorylation in your experimental model system, you can verify pathway activation in your cell lines without spending excess time and effort performing an analysis of immunoprecipitation and/or Western Blot. To use the phospho-array, treated or untreated cell lysate is added into antibody array glass slide wells. The subarray wells are washed, and biotinylated anti-phospho Tyr/Thr/Ser antibodies are then used to detect the phosphorylated Tyrosine/Threonine/Serine on target proteins. After incubation with a fluorescent dye-conjugated streptavidin (Cy3 equivalent), the slides can then be imaged using a laser scanner, such as the Axon GenePix, using the Cy3 channel.
Product Features
- High sensitivity and specificity
- Low sample volume (10-100 µl per array)
- Large dynamic range of detection
- Compatible with most sample types
- Test 4 or 8 samples on each slide
- Suitable for high-throughput assays
Target Names
6Ckine, Activin A, Activin B, Activin C, Activin RIA (ALK-2), Activin RIB (ALK-4), Activin RII A/B, Activin RIIA, Adiponectin (Acrp30), AgRP, ALCAM, Angiogenin, Angiopoietin-1, Angiopoietin-2, Angiopoietin-4, Angiopoietin-like 1, Angiopoietin-like 2, Angiopoietin-like Factor, Angiostatin, APJ, APRIL, AR (Amphiregulin), Artemin, Axl, B7-1 (CD80), BAFF R (TNFRSF13C), BCMA (TNFRSF17), BD-1, BDNF, beta-Catenin, BAX, beta-NGF, BIK, BLC (BCA-1/CXCL13), BMP-2, BMP-3, BMP-3b (GDF-10), BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-15, BMPR-IA (ALK-3), BMPR-IB (ALK-6), BMPR-II, BTC, Cardiotrophin-1 (CT-1), CCL14 (HCC-1/HCC-3), CCL28 (VIC), CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CD14, CD27 (TNFRSF7), CD30 (TNFRSF8), CD30 Ligand (TNFSF8), CD40 (TNFRSF5), CD40 Ligand (TNFSF5/CD154), CD 163, Cerberus 1, Chem R23, Chordin-Like 1, Chordin-Like 2, CLC, CNTF, CNTF R alpha, Coagulation Factor III (Tissue Factor), CRIM 1, Cripto-1, CRTH-2, Cryptic, Csk, CTACK (CCL27), CTGF (CCN2), CTLA-4 (CD152), CV-2 (Crossveinless-2), CXCL14 (BRAK), CXCL16, CXCR1 (IL-8 RA), CXCR2 (IL-8 RB), CXCR3, CXCR4 (fusin), CXCR5 (BLR-1), CXCR6, D6, DAN, DANCE, DcR3 (TNFRSF6B), Decorin, Dkk-1, Dkk-3, Dkk-4, DR3 (TNFRSF25), DR6 (TNFRSF21), Dtk, EDA-A2, EDAR, EDG-1, EGF, EGF R (ErbB1), EG-VEGF (PK1), EMAP-II, ENA-78, Endocan, Endoglin (CD105), Endostatin, Endothelin, EN-RAGE, Eotaxin (CCL11), Eotaxin-2 (MPIF-2), Eotaxin-3 (CCL26), Epiregulin, ErbB2, ErbB3, ErbB4, Erythropoietin, E-Selectin, FADD, FAM3B, Fas (TNFRSF6), Fas Ligand, FGF Basic, FGF-BP, FGF R3, FGF R4, FGF R5, FGF-4, FGF-5, FGF-6, FGF-7 (KGF), FGF-8, FGF-9, FGF-10 (KGF-2), FGF-11, FGF-12, FGF-13 1B, FGF-16, FGF-17, FGF-18, FGF-19, FGF-20, FGF-21, FGF-23, FLRG, Flt-3 Ligand, Follistatin, Follistatin-like 1, Fractalkine, Frizzled-1, Frizzled-3, Frizzled-4, Frizzled-5, Frizzled-6, Frizzled-7, Galectin-3, GASP-1 (WFIKKNRP), GASP-2 (WFIKKN), GCP-2 (CXCL6), GCSF, G-CSF R (CD 114), GDF1, GDF3, GDF5, GDF8, GDF9, GDF11, GDF-15, GDNF, GFR alpha-1, GFR alpha-2, GFR alpha-3, GFR alpha-4, GITR (TNFRF18), GITR Ligand (TNFSF18), Glucagon, Glut1, Glut2, Glut3, Glut5, Glypican 3, Glypican 5, GM-CSF, GM-CSF R alpha, Granzyme A, GREMLIN, GRO, GRO-a, Growth Hormone (GH), Growth Hormone R (GHR), HB-EGF, HCC-4 (CCL16), HCR (CRAM-A/B), Hepassocin, GLO-1, HGF, HGFR, HRG-alpha, HRG-beta 1, HVEM (TNFRSF14), I-309, ICAM-1, ICAM-2, ICAM-3 (CD50), ICAM-5, IFN-alpha/beta R1, IFN-alpha/beta R2, IFN-beta, IFN-gamma, IFN-gamma R1, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4, IGFBP-6, IGFBP-rp1 (IGFBP-7), IGF-I, IGF-I SR, IGF-II, IGF-II R, IL-1 alpha, IL-1 beta, IL-1 F5 (FIL1delta), IL-1 F6 (FIL1 epsilon), IL-1 F7 (FIL1 zeta), IL-1 F8 (FIL1 eta), IL-1 F9 (IL-1 H1), IL-1 F10 (IL-1HY2), IL-1 R3 (IL-1 R AcP), IL-1 R4 (ST2), IL-1 R6 (IL-1 Rrp2), IL-1 R8, IL-1 R9, IL-1 ra, IL-1 sRI, IL-1 sRII, IL-2, IL-2 R alpha, IL-2 R beta (CD122), IL-2 R gamma, IL-3, IL-3 R alpha, IL-4, IL-4 R, IL-5, IL-5 R alpha, IL-6, IL-6 R, IL-7, IL-7 R alpha, IL-8, IL-9, IL-10, IL-10 R alpha, IL-10 R beta, IL-11, IL-12 p40, IL-12 p70, IL-12 R beta 1, IL-12 R beta 2, IL-13, IL-13 R alpha 1, IL-13 R alpha 2, IL-15, IL-15 R alpha, IL-16, IL-17, IL-17B, IL-17B R, IL-17C, IL-17D, IL-17E, IL-17F, IL-17R, IL-17RC, IL-17RD, IL-18 BPa, IL-18 R alpha (IL-1 R5), IL-18 R beta (AcPL), IL-19, IL-20, IL-20 R alpha, IL-20 R beta, IL-21, IL-21 R, IL-22, IL-22 BP, IL-22 R, IL-23, IL-23 R, IL-24, IL-26, IL-27, IL-28A, IL-29, IL-31, IL-31 RA, BACE-1, FACX, Insulin, Insulin R, Insulysin (IDE), IP-10, I-TAC (CXCL11), Kininostatin (kininogen), Kremen-1, Kremen-2, Latent TGF-beta bp1, LBP, Lck, LECT2, Lefty - A, Leptin (OB), Leptin R, LFA-1 alpha, LIF, LIF R alpha, LIGHT (TNFSF14), Lipocalin-1, Lipocalin-2, LRP-1, LRP-6, L-Selectin (CD62L), Lymphotactin (XCL1), Lymphotoxin beta (TNFSF3), Lymphotoxin beta R (TNFRSF3), MAC-1, MCP-1, MCP-2, MCP-3, MCP-4 (CCL13), M-CSF, M-CSF R, MDC, MFG-E8, MFRP, MICA, MIF, MIG, MIP-1a, MIP-1b, MIP-1d, MIP 2, MIP-3 alpha, MIP-3 beta, MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11 (Stromelysin-3), MMP-12, MMP-13, MMP-14, MMP-15, MMP-16 (MT3-MMP), MMP-19, MMP-20, MMP-24 (MT5-MMP), MMP-25 (MT6-MMP), MSP alpha Chain, Musk, NAP-2, NCAM-1 (CD56), Neuritin, NeuroD1, Neuropilin-2, Neurturin, NGF R, Nidgen-1, NOV (CCN3), NrCAM, NRG1 Isoform GGF2, NRG2, NRG3, NT-3, NT-4, Orexin A, Orexin B, OSM, Osteoactivin (GPNMB), Osteocrin, Osteoprotegerin (TNFRSF11B), OX40 Ligand (TNFSF4), PARC (CCL18), PD-ECGF, PDGF R alpha, PDGF R beta, PDGF-AA, PDGF-AB, PDGF-BB, PDGF-C, PDGF-D, PECAM-1 (CD31), Pentraxin3 (TSG-14), Persephin, PF4 (CXCL4), PlGF, PLUNC, Pref-1, Progranulin, Prolactin, P-selectin, RAGE, RANK (TNFRSF11A), RANTES, RELM beta, RELT (TNFRSF19L), ROBO4, S100 A8/A9, S100A10, SAA, SCF, SCF R (CD117), SDF-1 (CXCL12), sFRP-1, sFRP-3, sFRP-4, sgp130, SIGIRR, Siglec-5 (CD170), Siglec-9, SLPI, Smad 1, Smad 4, Smad 5, Smad 7, Smad 8, Prdx6, Soggy-1, Sonic Hedgehog (Shh N-terminal), SPARC, Spinesin, TACI (TNFRSF13B), Tarc, TCCR (WSX-1), TECK (CCL25), TFPI, TGF-alpha, TGF-beta 1, TGF-beta 2, TGF-beta 3, TGF-beta 5, TGF-beta RI (ALK-5), TGF-beta RII, Grb2, TGF-beta RIII, Thrombopoietin (TPO), TPX, Thrombospondin-1, Thrombospondin-2, Thrombospondin-4, Thymopoietin, Tie-1, Tie-2, TIMP-1, TIMP-2, TIMP-3, TIMP-4, TL1A (TNFSF15), TLR1, TLR2, TLR3, TLR4, TMEFF1 (Tomoregulin-1), TMEFF2, TNF-alpha, TNF-beta, TNF RI (TNFRSF1A), TNF RII (TNFRSF1B), TRADD, TRAIL (TNFSF10), TRAIL R1 (DR4/TNFRSF10A), TRAIL R2 (DR5/TNFRSF10B), TRAIL R3 (TNFRSF10C), TRAIL R4 (TNFRSF10D), TRANCE, TREM-1, TROY (TNFRSF19), TSG-6, TSLP R, TWEAK (TNFSF12), TWEAK R (TNFRSF12), Ubiquitin+1, uPA, uPAR, Vasorin, VCAM-1 (CD106), VE-Cadherin, VEGF, VEGF R2 (KDR), VEGF R3, VEGF-B, VEGF-C, VEGF-D, VEGI (TNFSF15), WIF-1, WISP-1 (CCN4), XEDAR
Application Notes
Suggested Application
Multiplexed Protein Detection; Detection of Relative Protein Expression; Detecting Patterns of Cytokine Expression; Biomarker Screening; Identifying Key Factors; Confirming a Biological Process
Kit Components
- Human Protein Phosphorylation Array G1 Slide(s)
- Blocking Buffer
- Biotinylated Anti-Phospho Tyr/Thr/Ser Antibodies
- Cy3 equivalent-Conjugated Streptavidin
- 20X Wash Buffer I Concentrate
- 20X Wash Buffer II Concentrate
- Wash Buffer III
- 2X Cell Lysis Buffer Concentrate
- Protease Inhibitor Cocktail
- 100x Phosphatase Inhibitor Cocktail I
- Phosphatase Inhibitor Cocktail II
- Adhesive film
Other Materials Required
- Shaker
- Laser scanner for fluorescence detection
- Aluminum foil
- Distilled water
- Plastic box
- 50 ml Centrifuge tube
- Isopropanol (2-propanol)
Protocol Outline
- Dry the Glass Slide
- Blocking and Incubation
- Fluorescence Detection
Storage/Stability
Upon receipt, the kit should be stored at ≤-20°C. Please use within 6 months from the date of shipment. After initial use, the 2X Cell Lysis Buffer, Blocking Buffer, 20X Wash Buffer I, 20X Wash Buffer II, Biotinylated Anti-Phospho Tyr/Thr/Ser Antibodies and Cy3 equivalent-Conjugated Streptavidin should be stored at 2-8°C to avoid repeated freeze-thaw cycles. The Array Glass Slide, Protease Inhibitor Cocktail and Phosphatase Inhibitor Cocktail Set II should be kept at ≤20°C. Use within 3 months after initial use.