Introduction
Protein S-Nitrosylation plays an unusually prominent role in cell signaling, development and growth. The RayBio® G-Series Human Protein S-Nitrosylation Antibody Array 2 is a very rapid, convenient, and sensitive assay that can simultaneously detect multiple protein S-Nitrosylation of cysteine and be used to monitor the activation or function of important biological pathways.
RayBiotech is committed to develop a series of S-Nitrosylation antibody arrays. This multiplex glass slide array is specifically designed for simultaneous identification of the relative levels of S-Nitrosylation of cysteine of 493 different human proteins in cell lysate, culture supernatant, serum, plasma and other biological samples. By monitoring the changes in protein S-Nitrosylation 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.
RayBio® S-Nitrosylation Detection Kit uses a modified 'biotin-switch' method (Jaffrey et al.) to allow for the direct visualization of S-nitrosylated proteins by western blot analysis as well as by antibody array. In this S-nitrosylation biotin switch assay, unmodified free cysteines are first blocked. S-nitrosylated cysteines are then selectively reduced for specific labeling with biotin-maleimide reagents, which irreversibly bind to the cysteine thiol that was S-nitrosylated. Biotinylation of the newly formed thiol groups can then be detected by western blot or antibody array. In addition, avidin resin can be used to selectively enrich S-nitrosylated proteins/peptides labeled with biotin. The biotin labeled sample is added into antibody array glass slide wells. The antibody array slide wells are washed. 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
11b-HSD1, 2B4, 4-1BB, A1BG, A2M, ABL1, ACE, ACE-2, ACK1, ACPP, ACTH, ADAM-9, ADAMTS-1, ADAMTS-10, ADAMTS-13, ADAMTS-15, ADAMTS-17, ADAMTS-18, ADAMTS-19, ADAMTS-4 , ADAMTS-5, ADAMTS-L2, Adipsin, Afamin, AFP, ALBUMIN, Aldolase A, Aldolase B, Aldolase C, ALK, Alpha 1 AG, Alpha 1 Microglobulin, Alpha Lactalbumin, ALPP, AMICA, AMPKa1, Amylin, ANGPTL3, ANGPTL4, Annexin A7, APC, APCS, Apelin, Apex1, APN, ApoA1, ApoA2, ApoA4, ApoB, ApoB100, ApoC1, ApoC2, ApoC3, ApoD, ApoE, ApoE3, ApoH, ApoM, APP, ASPH, Attractin, B3GNT1, BAF57, BAFF, BAI-1, BCAM, Beta 2M, Beta Defensin 4, Beta IG-H3, Biglycan, BLAME, BMP-9, BMX, BNIP2, BNP, Btk, C2, C3a, C5/C5a, C7, C8B, C9, CA125, CA15-3, CA19-9, CA9, Cadherin-13, Calbindin, Calbindin D, Calcitonin, Calreticulin, Calsyntenin-1, CART, Caspase-3, Caspase-8, Cathepsin B, Cathepsin D, Cathepsin L, Cathepsin S, CBP, CCK, CD200, CD23, CD24, CD36, CD38, CD44, CD45, CD46, CD47, CD55, CD59, CD61, CD71, CD74, CD79 alpha, CD90, CD97, CEA, CEACAM-1, Ceruloplasmin, CFHR2, Chemerin, CHI3L1, Chromogranin A, Chymase, cIAP-2, Ck beta 8-1, CK-MB, Claudin-3, Claudin-4, CLEC3B, Clusterin, CNDP1, COCO, Complement factor H, Contactin-1, Contactin-2, Corticosteroid-binding globulin, COX-2, C-peptide, CPN2, Creatinine, CRP, CRTAM, CSH1, cTnT / Troponin T, CutA, Cyclin D1, Cystatin A, Cystatin B, Cystatin C, Cytochrome C, Cytokeratin 8, Cytokeratin18, Cytokeratin19, DBI, DCBLD2, D-Dimer, DEFA1/3, Defensin, Desmin, DLL1, DLL4, DMP-1, DPPIV, E-Cadherin, Endorphin Beta, Endothelin Receptor A, Enolase 2, ENPP2, EpCAM, EphA1, EphA2, EphA3, EphA4, EphA5, EphA6, EphA7, EphA8, EphB1, EphB2, EphB3, EphB4, EphB6, ERRa, Erythropoietin R, ESAM, EV15L, EXTL2, FABP1, FABP2, FABP3, FABP4, Factor XIII A, Factor XIII B, FAK, FAP, Fc RIIB/C, Fen 1, FER, Ferritin, Fetuin A, Fetuin B, FGFR1, FGFR1 alpha, FGFR2, Fibrinogen, Fibrinopeptide A, Fibronectin, Ficolin-3, FIH, FOLR1, FOXN3, FoxO1, FoxP3, FRK, FSH, Furin, Fyn, GADD45A, Galanin, Galectin-1, Galectin-3BP, Galectin-7, gamma-Thrombin, Gas1, Gastrin, GATA-3, GATA-4, Gelsolin, Ghrelin, GLP-1, GMNN, GPBB, GPI, GPR-39, GPX1, GPX3, GRP, GRP75, GRP78, GSR, GST, HADHA, HAI-1, HAI-2, Haptoglobin, hCG alpha, hCGb, Hck, HE4, Hemopexin, Hepcidin, HOXA10, HSP10, HSP20, HSP27, HSP32, HSP40, HSP60, HSP70, HSP90, HSPA8, HTRA2, IBSP, IGF2BP1, IGFBP-5, IL-23p19, IL-33, IL-34, IL36RN, INSL3, INSRR, Integrin alpha V, Itk, ITM2B, Kallikrein 10, Kallikrein 11, Kallikrein 14, Kallikrein 2, Kallikrein 5, Kallikrein 6, Kallikrein 7, Kallikrein 8, KCC3, KCTD10, KIF3B, KLF4, LAG-3, Layilin, LDL R, Legumain, LH, LIMPII, LIN41, Livin, LOX-1, LPS, LRG1, LTF, LTK, Lumican, Lyn, LYRIC, LYVE-1, LZTS1, Mammaglobin A, Marapsin, MATK, MBL, MBL-2, Mer, Mesothelin, MICB, Midkine, MINA, MSHa, MTUS1, Myoglobin, NAIP, Nanog, NELL2, NEP, Nesfatin, Nestin, NET1, Netrin G2, Netrin-4, Neurokinin-A, Neuropeptide Y, NF1, NM23-H1/H2, Notch-1, NPTX1, NPTXR, NR3C3, Ntn1, OCT3/4, Omentin, Osteocalcin, Osteopontin, OX40, p21, p27, p53, PAI-1, PAK7, Pancreastatin, Pancreatic Polypeptide, Pappalysin-1, PARK7, P-Cadherin, PCAF, PD-1, PDX-1, PEDF , PEPSINOGEN I, PEPSINOGEN II, PGRP-S, PI 16, PI 3Kinase p85 beta, PIM2, PKM2, Plasminogen, Podocalyxin, POMC, PON1, PON2, PPARg2, PPP2R5C, Presenilin 1, Presenilin 2, Pro-BDNF, Procalcitonin, Pro-Cathepsin B, Progesterone, pro-Glucagon, Prohibitin, Pro-MMP-13, Pro-MMP-7, Pro-MMP-9, ProSAAS, Prostasin, Protein p65, PSA-Free, PSA-total, PSP, PTH, PTHLP, PTN, PTPRD, PYK2, PYY, Ras, RBP4, RECK, RELM alpha, Resistin, RET, RIP1, ROCK1, ROCK2, ROR1, ROR2, ROS, RYK, S100A4, S100A6, S100A8, S-100b, SART1, SART3, SCG3, Selenoprotein P, SEMA3A, Serotonin, Serpin A1, Serpin A12, Serpin A3, Serpin A4, Serpin A5, Serpin A8, Serpin A9, Serpin B5, Serpin D1, Serpin I1, SERTAD2, SHBG, SMAC, SNCG, Somatotropin, SOST, SOX17, SOX2, SPARCL1, SPINK1, SRMS, SSEA-1, SSEA-4, SSTR2, SSTR5, Survivin, SYK, Syndecan-1, Syndecan-3, TACE, TAF4, Tec, TFF1, TFF3, Thrombin, Thrombomodulin, Thymidine Kinase-1, Thyroglobulin, TIM-1, TNK1, TOPORS, TPA, TPM1, TRA-1-60, TRA-1-81, Transferrin, Trappin-2, TRKB, Troponin C, Troponin I, TRPC1, TRPC6, TRPM7, Trypsin 1, TSH, TSLP, TXK, Tyk2, TYRO10, Uromodulin, Vasopressin, VDUP-1, VEGF R1, VGF, VIP Receptor 2, Visfatin, Vitamin D Receptor, Vitamin D-BP, Vitamin K-dependent protein S, Vitronectin, VWF, Wilms Tumor 1, XIAP, ZAG, ZAP70
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
- RayBio® Glass Slide(s)
- Blocking Buffer
- 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
- S-Nitrosylation Buffer A
- S-Nitrosylation Buffer B
- S-Nitrosylation Buffer C
- S-Nitrosylation Buffer D
- S-Nitrosylation Buffer E
- S-Nitrosylation Blocking Reagent
- S-Nitrosylation Reduction Reagent I
- S-Nitrosylation Reduction Reagent II
- S-Nitrosylation Labeling Reagent
- Adhesive film
Other Materials Required
- Acetone, ≥ 98% (hazardous)
- 1.5 mL microcentrifuge tubes
- 15 mL tubes (polypropylene)
- 10 mL graduated cylinders (X2)
- Benchtop centrifuge and microcentrifuge (4°C)
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- Water bath or heat block
- 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.
