Human Glycosylation Array L3

Introduction Recent technological advances have enabled the largest commercially available glycosylation antibody array to date. With the Human Glycosylation Array L3, researchers can now efficiently obtain glycosylation profiles in their samples. The glycosylation profiles of 500 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 14-3-3 beta, 14-3-3 epsilon, 14-3-3 eta, 14-3-3 gamma, 14-3-3 sigma, 14-3-3 theta, 14-3-3 zeta, 53BP1, LAMR1, ABAT, ABCF1, ABI3BP, ACAA1, ACAA2, ACACA, ACAA, ACLP, ACLY, Aconitase 1, ACTBL2, ACTC1, Actinin alpha 1, ADAMDEC1, ADAS, ADH1B, ADH1C, ADH4, ADH5, ADM, Advillin-N-t, AFG3L2, AGA, Aggrecan, AGXT, AHNAK , Ahsp, AIF, AK2 , AKAP9 , AKR1B1, AKR1C3, AKR7A2, ALAD, Alanine Transaminase, Alcohol Dehydrogenase, Aldehyde Oxidase 1, ALDH16A1 , ALDH1A1, ALDH9A1, Alkaline Phosphatase, ALP, alpha 1,2 Mannosidase IA, alpha Actinin 4, Alpha Fodrin, alpha Glucosidase II, Alpha-synuclein, alpha Tubulin, AlphaA Crystallin, ALS, Als2 , ALS+B62:B1212CR1 , Aminoacylase-1, Androgen Receptor, ANGPTL6, ANGPTL8, ANK, Ankrd26, Annexin A1, Annexin A2, Annexin A6, Annexin V, ANP, Antithrombin 3, APA, APLP-1, APM2, Apo (a), APOA1BP, ApoF, ApoL1, ApoL2, ARFBP1, ARFGEF3, Argininosuccinate Lyase, ArgRS, ARP19, Arp2, ARP2/3, Arp3, ARPC2, ARPC3, ART3, Aminopeptidase PILS, ARX, ASH2L , Asialoglycoprotein Receptor 2, ASK1 , Aspartate Aminotransferase , Aspartyl Aminopeptidase, ASXL1, ATBF1, ATP5A, ATP5O, ATPB, B3GNT2, B4GalT1, B7-H2, B7-H3, bad, Band 3, BASP1, Bassoon, BAZ2B, BCHE, bcl-w, BCOR, beta 1 Spectrin , beta B1 Crystallin, Tubulin beta-1 chain, Tubulin beta-3 chain, BID, BIN2, BIRC6, BLMH, BLVRB, BMP-1, BPGM, BPIFB1, BPIL1, BRCA 2, BRD2, Brevican, Brg1, BRSK1, BTD, BTF3, C1Q, C1QA, C1QB, C1QR1, C1RL, C1S, C3orf75, C4.4A, C4BPA, C5b-9, C6 -N-terminal, C8G, C9orf40, CA1, CA150 , CA2, CA3, CACNB4, CAD , Cadherin-22, Cadherin-6, Caldesmon, CALML5, Calmodulin, Calpain 1, Calpain S1 , Calpastatin, Calretinin, Calumenin , CAP1, CapG, CAPZA1, Carboxypeptidase B2, CARHSP1, Caspase-14, Catalase, Cathelicidin, Cathepsin A, Cathepsin G, Cathepsin H, Cathepsin Z, CBS, CCDC126, CCDC25, CCT3, CD109, CD133, CD155, ADP-Ribosyl Cyclase 2, CD16, CD21, CD32, CD35, CD39L4, CD41, CD42b, CD48, CD5L, CD9 , CD98, CDA, CDC5L, CDK2 , CEACAM-8, CECR1, CENPF, CEP57, Carboxylesterase 1, CETP, Cezanne, CFHR 1, CFHR4, Complement Factor H-related 5, CFI, CFL1, Coagulation Factor VII, CHC17, Chitobiase, Chitotriosidase, CHORDC1, CHREBP, Chromogranin B, Chromogranin C, CIP29, CKB, CLIC1, CLIC4, CLIP170-N-t, Collectin-12, CLPS, CLTA, CNN2, CNOT1, CO4A2, COG4 , COL19A1, COL4A3, COL6A2, COL9A3, COLEC10, Collagen I alpha-1, Collagen III, Collagen IV alpha-6, Collagen IX, Collagen V, Collagen VI, Collagen X, Collagen XV alpha 1, Thrombospondin 5, Complement Factor B, Contactin-3, COPS8 , Corneodesmosin, Coronin 3, Cortactin, COTL1, Carboxypeptidase E, CPEB3, Carboxypeptidase M, CPN1, CPNE3, CPS1, Creatine Kinase MM, CRF21, CRHBP, CrkL, CRMP2, CRTAC1, CS, Ctip2, Cux2, Cyclophilin A, Cyclophilin B, Cystatin D, Cystatin E/M, Cystatin S, Cystatin SN, Cysteine-rich Protein 1, CYTL1, Cytochrome b5, Cytochrome C, Cytokeratin 1, Cytokeratin 10, Cytokeratin 13, Cytokeratin 14, Cytokeratin 15, Cytokeratin 16, Cytokeratin 17, Cytokeratin 20, Cytokeratin 3, Cytokeratin 4, Cytokeratin 5, Cytokeratin 9, D4-GDI, DAK, DAN, DARS2, DCI, DCXR, DDAH1, DDT, DDX3Y, DEFA6, DEP-1, Der p2, Dermcidin, Desmocollin-1, Desmocollin-2, Desmocollin-3, Desmoglein-1, Desmoglein-2, Desmoplakin, Desmuslin, Destrin , DGK, DISC 1, DMGDH, DMRN9, Dopamine beta Hydroxylase, DOT1L, DPEP2, DPP3, DPPI, DRIL1, DSCAM, DSPG3, Dystroglycan, Ubiquitin-activating enzyme E1, ECHS1 , ECM-1, EEF1G , EEF2, EFEMP2 , EFTUD2, EHD1 , EHD3, EIF3S2, eIF4A1-N-terminal, eIF5A, ELAVL1 , EMILIN1 , EMSY, EN2, Endorepellin, ENO1, ENO1/2/3, ENSA, Envoplakin, Eosinophil derived neurotoxin, EPB41, EPCR, Ephrin-B1, Ephrin-B2, EPHX2, EPPK1, Eps 15, ERAB, Aminopeptidase LRAP, ERp29, ERp57, ERp72, ESD, ESR1, ETL, EVC2, Ezrin, Coagulation Factor XI, FABP5, Coagulation Factor IX, Coagulation Factor V, Coagulation Factor XII, Coagulation Factor XIII, FAM20C, FAM3C, Fascin, FASN, fast skeletal Myosin, FASTKD5, F-box protein 38, FBPase 2, FBPase 1, FCGBP, FDPS , FH, Fibrillin 1, Fibrinogen gamma chain, Fibrinogen-like 2, Fibrinopeptide B, Fibulin 3, Ficolin-2, Filamin A, Filamin B, Filamin C, FKBP12, FKBP25, FKBP51, FLG2, FOLR3, Frizzled 8, FRY, FSH, FTL, FUCA1, FUCA2, Fumarylacetoacetate hydrolase, G0/G1switch 2, G3BP , GALNT2, Catenin gamma, GAPHD, GARNL1 , GART, Gastrokine 1, GATM C-terminal, GBE1, GCDFP15, GCLC , GCSH , GDA, GDF7, GDI1, GDI2, Gephyrin, GFAP, GHRF, GIP, GLIPR2, GLRX1, G6PD, Glucosidase 2 subunit beta, GLUD1, Glutamyl hydrolase gamma , GSTO1, Glutathione Synthetase, Glycerol 3 Phosphate Dehydrogenase, GPV, Glyoxalase II, GM2A, GMF beta, GNB1, GNPTG, GOLM1, GOLPH4 , GOT2, GPCR GPR116, GPLD1, Grainyhead-like protein 1 homolog, Granzyme M, GRHPR, GRP, GSTM1, GSTP1, Guanylin, GULP1, H6PD, HABP2, HBZ, HCFC1, HDGF, HEG1, Hemoglobin, Hemoglobin A1c, Hemoglobin subunit beta, Hemoglobin subunit delta, Hemoglobin subunit gamma 2, HEXB, HGFA, Growth Hormone, hHR23b, HIBADH , HINT1, HIP1R , Histone H1.2, Histone H1.3, Histone H2A, Histone H2A.Z, Histone H2B K, Histone H3.3, Histone H4, HLA-C, HMGB1, HMGB2, HMGB3, HMGN2, HN1, HNF-3 alpha , hnRNP A1, hnRNP A2B1, hnRNP C1/C2, hnRNP G , hnRNP L, hnRNP M1-M4 , hnRNP U , Hornerin, HOXB3, HOXD11, HP1BP3, HPD, HPR, HPRT, HPRG, HRSP12, HSC70, HSP47 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 L3 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 L3 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 500 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.