Introduction
Protein S-acylation (or protein S-palmitoylation) plays an unusually prominent role in cell signaling, development and growth. By monitoring the changes in protein S-Acylation 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.
The array kit uses a modified 'biotin-switch' (acyl-biotinyl exchange (ABE)) method (Jaffrey et al.) to allow for the direct visualization of S-acylated proteins on the antibody array. Prior to running the array, the samples are labeled. Unmodified free cysteines on proteins in the sample are first blocked. S-acylated cysteines are then selectively reduced for specific labeling with biotin-maleimide reagents, which irreversibly bind to the cysteine thiol that was S-acylated. Biotinylation of the newly formed thiol groups can then be detected on the antibody array. If desired, avidin resin can be used to selectively enrich S-acylated 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-Acylation Buffer A
- S-Acylation Buffer B
- S-Acylation Buffer C
- S-Acylation Buffer D
- S-Acylation Buffer E
- S-Acylation Blocking Reagent
- S-Acylation Reduction Reagent
- S-Acylation 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.
