Human Glycosylation Array L4

Introduction Recent technological advances have enabled the largest commercially available glycosylation antibody array to date. With the Human Glycosylation Array L4, 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 HSPA1A, HTRA1, Human Agrin, IBP160 , IDH1, IDH3A, IFRD1 , IMP2, IGFBP-rp1, IGSF4B, Ihh, ILK, Inhibin beta, Integrin b1, Integrin beta 6, Intergrin a6, IQGAP1 , IQGAP2 , IRE1, IRS2, ISOC2, ITGB4BP, ITIH1, ITIH2, ITIH3, ITIH4 a, JAM-A, JARID2, Karyopherin beta 1, Keratin 36, Keratin 38, KHSRP, KIAA0319L, KIAA1468, KIAA1967, KIF5B , Kilon, KLK-B1, KMD4B, KMT2B, KRT31, KRT72, KRT73, KRT82, KRT85 N-terminal, KRTDAP, KRTHA3B, KSR1, LAD, LAF4, LAIR1, LAM b1, LAMA, Lamin A/C, Lamin B1, Lamin B2, Laminin 2 alpha, Laminin b2, Laminin gamma 1, LAMP, LAMP1, LAMP2, LAP3, LASP1, Latent TGF beta bp2, LCAT, LCMT2, LDHA, LDHB, LEDGF, LEKTI, LILRA3, LIMS1, LMAN2, LMW-PTP, LOK, LOX, LOXL1, LRP 4, LTA4H, LTBP4, Lubricin, LUZP1, LYPA1, Lysozyme, MAGI2, MAGP-2, MAN1, MANF, Mannosidase II, MAP1A, MAPRE1, MARCKS, Complement MASP3, MBD2 , MBP, MCAM, Mcl-1, MCM, MCM5 , MCMP2, MDH1, MDH2, ME1, Meprin alpha, Metallothionein, Metavinculin, MFAP4, MFI2, mGLUR5, MGP, Mimecan, MINPP1, MLCK, MMR, MMRN1, MN1, Moesin, MP1, MPCA, Myeloperoxidase, MRP 1, MSH6, mTOR, MUCDHL, Multimerin 2, MyBPC3, MYH2, MYH6, MYH7, MYHC, MYL12B , MYL3, MYO5A, Myoferlin , Myosin 18B, Myotrophin, NABC1, NAGLU, NAP1L1 , NAPRT1, NASP , NCAM2 , Nebulin, Nectin-1, Nectin-3, NEDD8, Neogenin, Nesprin2, Neurabin 1, N-Cadherin, Neurofibromin, Neurogranin, Neuropeptide B, Neuropilin-1, Neurotrimin, NF-M, Nidogen-2, NIT2, NME3, nNOS, Noelin, Non-muscle Actin, Non-muscle Myosin IIA, Notch-2, Notch-2 ICD, NPAS3, NPM1, NQO2, NT5C3, Nucleobindin 1, NUP98, OBCAM, OIT3, Olfactomedin-2, Ornithine Carbamoyltransferase , Orosomucoid 2, ORP150, OSBP1, OSCAR, Oncostatin M R beta, Osteoadherin, Oxytocin-neurophysin 1, p16 ARC , P20Sb3, p23, p39 , P4HB, p73, PA2G4, PABP, PACS1, PARVB, PCBP1, PCBP2, PCCA, PCDH7, PCDX8, PCK2 , PCMT1, PCNA, PCPE-1, Proprotein Convertase 9, PCYOX1, PDE1B, PDIA6, PDLIM1, PDLIM5, PDZD2, PEBP4, PEPD, PER1, Perilipin-3, Perilipin-1 , Periostin, Peroxiredoxin 2, Peroxiredoxin 3, Peroxiredoxin 5, PF4V1, PGAM1, PGAM2, PGD, PGDF, PGK-1, PGLS-C-t, PGM1, PGRPL, PHAP1, Phosphoserine Aminotransferase, PI3-Kinase C2 beta, pIgR, PIK3IP1, PIN, PISD, PKLR, PLA2G1B, Plakophilin 1, Plastin L, PLC-gamma 1 , Pleckstrin, Plectin, Plexin B1, Plexin B2, PLOD1, PLOD2, PLS3, Plxdc2, PNP, POR, PPCS, PPOX , PPP2R1B , PPP2R4, PRCP, PRDM13, PRDX 1, PRELP, PREP, PRG2, Prion protein PrP , Profilin 1, Properdin, Prosaposin, Prostaglandin D Synthase, Proteasome 20S a+b, Proteasome 20S alpha, Proteasome 20S alpha 5, Proteasome 20S b7, Proteasome 26S S5, Proteasome beta 1, Proteasome subunit alpha type 6, Proteasome subunit beta type 2, Proteasome subunit beta type 4, Coagulation Factor XIV, Protein Z, Prouroguanylin, PRSS23, PRSS3, Myeloblastin, PSMA1, PSMA2, PSMA4, PSMA7, PSMB5, PSMC3 , PSMD1, PSMD9 , PTEN, PTK 7, PTMA, PTP gamma, PTP kappa, PTP mu, PTPRS, PTPRZ, PYGL, PZP, QDPR, QPRT, Quiescin Q6, Rab7a, Ran, RanGAP1, RAP1AB, Rbm15, RCL , Reg1A, Reg3A, RHOC, RhoGDI, Ribonuclease A, Ribonuclease Inhibitor, Ribonuclease T2, RKIP, RNA Polymerase II, RNASE4, RNASE6, RPL10, RPL10A , RPL11, RPL12, RPL14 , RPL17, RPL22, RPL5, RPL7A, RPLP0 , RPS10, RPS11, RPS12, RPS19, RPS2, RPS20 , RPS23, RPS25, RPS28, RPS3, RPS5, RREB1, RSU1, S100 A1, S100 A11, S100 A7, S100 A9, S100P, SAA, SAA4a, Salivary alpha amylase, SAMSN1, SBP-1, SBSN, SDF4, SDNSF, SDPR, Secretogranin V, Semaphorin 6B, Semaphorin 7A, Semenogelin I, Semenogelin II, Serpin A11, Serpin A7, Serpin B3, Serpin B6, Serpin B8, Serpin F2, Serpin A10, Serpin B1, Serpin B4, Serpin E2, SerRS, SET, SEZ6L2, SF20, SH3BGRL, SH3BGRL3, SHANK1, SHC1, SHIP, SHMT1, SHP-1, Siglec-1, Siglec-14, SIM2 , SIRP beta 1, Six3, SLC38A10, SLITRK1, SLURP1, SMA, SMC4 , SMPD4, SOD1, SOD2, SOD3, SOD4, Somatostatin, SORD, SorLA, SOX4, SP-D, Spectrin beta-5, SPEN, SPINK7, SPTBN1, Src(1), SREC-II, STAT3, Stathmin 1, Sterol carrier protein 2, STI1, STOM, SUCLG1, SUMO3, Symplekin, SynCAM, Syntaxin 7, TAB182, TAGLN2, Talin1, Talin1&2, TAX1BP3 , TBCA, TCEB2, Tcf20, TCN1, TCP1 eta, Tenascin C, Tenascin X(1), TFF2(1), TGM3, Thioredoxin-1, Thimet Oligopeptidase, Thymosin b10, Titin, TLS, TMEM223, TOB2 , TOP2B, TPM4, Tripeptidyl-peptidase I, Transaldolase 1, Transketolase, Transthyretin, TRAP1, TRAP220, TRF 2, Triosephosphate isomerase, Tropomyosin 3, TRP-1, TRPS1, Trypsinogeb-2, TrypsinPan, Tryptophanyl, TSR2, TUBA6, TWF2, TXNDC4, TXNDC5, TXNRD2, UBE2D3, Ube2L3, UBE2N, Ubiquitin+1, UCH-L1, UFM 1, UGGT, UNC-13 Homolog D, UNC45A, UNC5H4, UPB1 , UQCRB, UQCRH , URB, URB2, UROC1, UROD, URP2, USP14 , USP5 , Uteroglobin, Utrophin, VAP-1, VAP-A, VCP, VDAC1 , Versican isoform V0, Vimentin B, VNN1, VSIG4, WDR1, WISP2, WNK2, YB1 , YY1, ZBTB4, ZC3H4-N-t, ZC3H8, ZDHHC18 , ZNF671, Zyxin 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 L4 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 L4 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 L4 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.