Human Glycosylation Array 507

Introduction Recent technological advances have enabled the largest commercially available glycosylation antibody array to date. With the Human Glycosylation Array 507, researchers can now efficiently obtain glycosylation profiles in their samples. The glycosylation profiles of 507 human proteins can be simultaneously detected, including, but not limited to, cytokines, chemokines, adipokines, growth factors, angiogenic factors, proteases, soluble receptors, soluble adhesion molecules in cell culture supernatants, serum, plasma and other samples types. Capture antibodies are printed onto glass slides and the glycans on these capture antibodies are removed. The glass slide arrays come pre-blocked and are ready to be incubated with samples. After incubation with samples and washing to remove unbound proteins, five unique biotin-labeled lectins are incubated with the array to allow binding to the glycans from captured target proteins. Streptavidin-conjugated fluorescent dye (Cy3 equivalent) is then added to the array. Finally, the glass slide is dried and laser fluorescence scanning is used to visualize the signals. These signals are then compared to the array map to identify glycosylated proteins present in the samples. Product Features High density screening High sensitivity and specificity Low sample volume (20-100 µl per array) Large dynamic range of detection Compatible with most sample types Accurate and reproducible Target Names 6Ckine, Activin A, Activin B, Activin C, Activin RIA (ALK-2), Activin RIB (ALK-4), Activin RII A/B, Activin RIIA, Adiponectin, AgRP, ALCAM, Angiogenin, Angiopoietin 1, Angiopoietin 2, Angiopoietin-4, ANGPTL1, ANGPTL2, Angiopoietin-like Factor, Angiostatin, APJ, APRIL, Amphiregulin, Artemin, Axl, B7-1 (CD80), BACE-1, BAFF R (TNFRSF13C), bax, BCMA (TNFRSF17), BD-1, BDNF, beta-Catenin, b-NGF, BIK, BLC (BCA-1, CXCL13), BMP-15, BMP-2, BMP-3, BMP-3b (GDF-10), BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, 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, CD 163, CD14, CD27 (TNFRSF7), CD30 (TNFRSF8), CD30 Ligand (TNFSF8), CD40 (TNFRSF5), CD40 Ligand (TNFSF5, CD154), Cerberus 1, Chem R23, Chordin-Like 1, Chordin-Like 2, CLC, CNTF, CNTF R alpha, Coagulation Factor III (Tissue Factor), Coagulation Factor X/Xa, CRIM 1, Cripto-1, Crossveinless-2 (CV-2), CRTH-2, Cryptic, Csk, CTACK (CCL27), CTGF (CCN2), CTLA-4 (CD152), 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, EGFR (ErbB1), EG-VEGF (PK1), EMAP-II, ENA-78, Endocan, Endoglin (CD105), Endostatin, Endothelin, EN-RAGE, Eotaxin-1 (CCL11), Eotaxin-2 (MPIF-2), Eotaxin-3 (CCL26), Epiregulin, ErbB2, ErbB3, ErbB4, Erythropoietin, E-Selectin, FADD, FAM3B, Fas (TNFRSF6), Fas Ligand, bFGF, FGF R3, FGF R4, FGF R5, FGF-10, FGF-11, FGF-12, FGF-13 1B, FGF-16, FGF-17, FGF-18, FGF-19, FGF-20, FGF-21, FGF-23, FGF-4, FGF-5, FGF-6, FGF-7 (KGF), FGF-8, FGF-9, FGF-BP, 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), G-CSF, G-CSF R (CD 114), GDF1, GDF11, GDF15, GDF3, GDF5, GDF8, GDF9, GDNF, GFR alpha-1, GFR alpha-2, GFR alpha-3, GFR alpha-4, GITR (TNFRF18), GITR Ligand (TNFSF18), GLO-1, Glucagon, Glut1, Glut2, Glut3, Glut5, Glypican 3, Glypican 5, GM-CSF, GM-CSF R alpha, Granzyme A, Grb2, GREMLIN, GRO, GRO alpha, Growth Hormone, Growth Hormone R, HB-EGF, HCC-4 (CCL16), HCR (CRAM-A/B), Hepassocin, 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-gammabeta, IFN-gamma, IFN-gamma R1, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4, IGFBP-6, IGFBP-rp1 (IGFBP-7), IGF-1, IGF-1 sR, IGF-2, IGF-2 R, IL-1 alpha, IL-1 beta, IL-1 F10 (IL-1HY2), 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 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-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-2, IL-2 R alpha, IL-2 R beta (CD122), IL-2 R gamma, 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-3, IL-3 R alpha, IL-31, IL-31 RA, 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, Insulin, Insulin R, Insulysin (IDE), IP-10, I-TAC (CXCL11), KGF-2, 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, 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, MIF, MIG, MIP 2, MIP-1 alpha, MIP-1 beta, MIP-1 delta, MIP-3 alpha, MIP-3 beta, MMP-1, MMP-10, MMP-11 (Stromelysin-3), MMP-12, MMP-13, MMP-14, MMP-15, MMP-16 (MT3-MMP), MMP-19, MMP-2, MMP-20, MMP-24 (MT5-MMP), MMP-25 (MT6-MMP), MMP-3, MMP-7, MMP-8, MMP-9, MSP alpha chain, Musk, NAP-2, NCAM-1 (CD56), Neuritin, NeuroD1, Neuropilin-2, Neurturin, NGF R, NOV (CCN3), NRG1 Isoform GGF2, NRG2, NRG3, NT-3, NT-4, Orexin A, Orexin B, OSM, Osteoactivin (GPNMB), Osteocrin, Osteoprotegerin (Osteoprotegerin), 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), Peroxiredoxin 6 (Prdx6), 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, Smad1, Smad4, Smad5, Smad7, Smad8, Soggy-1, Sonic Hedgehog (ShhN-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, TGF beta RIII, Thrombopoietin (TPO), Thrombospondin-1, Thrombospondin-2, Thrombospondin-4, Thymopoietin, Thyroid Peroxidase (TPX), Tie-1, Tie-2, TIMP-1, TIMP-2, TIMP-3, TIMP-4, TL1A (TNFSF15), TLR1, TLR2, TLR3, TLR4, TMEFF1 (Tomoregulin-1), TMEFF2, TNF RI (TNFRSF1A), TNF RII (TNFRSF1B), TNF alpha, TNF beta, TNFSF14, 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-A, VEGFR2 (KDR), VEGFR3, VEGF-B, VEGF-C, VEGF-D, VEGI (TNFSF15), WIF-1, WISP-1 (CCN4), XEDAR Biotin-Labeled Lectin Mixture Lectin Name Sugar specificity 1 Concanavalin A αMan, αGlc 2 Dolichos Biflorus Agglutinin αGalNAc 3 Peanut Agglutinin Galβ3GalNAc 4 Ulex Europaeus Lectin 1 αFuc 5 Wheat Germ Agglutinin GlcNAc Application Data/Notes Typical Results The following figure shows the Human Glycosylation Antibody Array 507 probed with conditioned media sample. The images were captured using a Axon GenePix laser scanner. The strong signals in row 20 and the upper left and lower right corners of each array are Positive Controls, which can be used to identify the orientation and help normalize the results between arrays. Suggested Applications Identify and profile the glycans in their samples Determine whether their biomarker of interest has glycan moieties Determine relative expression levels Kit Components Human Glycosylation Array 507 Glass Slide/s Sample Diluent 20X Wash Buffer I 20X Wash Buffer II Cy3 equivalent dye-conjugated Streptavidin Biotin-labeled Lectin Mixture Adhesive device sealer Manual Other Materials Required Pipettors, pipet tips and other common lab consumables Orbital shaker or oscillating rocker Aluminum foil Distilled or de-ionized water 1 ml microcentrifuge tubes Gene microarray scanner or similar laser fluorescence scanner Protocol Outline Dry the glass slide Incubate samples Incubate with biotin lectin mixture Incubate with Cy3 Equivalent Dye-Streptavidin Disassemble the glass slide Scan with a  gene microarray laser scanner Perform densitometry and analysis Storage/Stability Upon receipt, all components of the Human Glycosylation Array 507 kit should be stored at -20°C. Once thawed, the glass slide and Cy3 equivalent dye-conjugated Streptavidin should be kept at –20°C and all other components may be stored at 4°C. The entire kit should be used within 6 months of purchase.