Protein S-Nitrosylation plays an unusually prominent role in cell signaling, development and growth. RayBio® Human Protein S-Nitrosylation Antibody Array 8000 is specifically designed for simultaneous identification of the relative levels of S-Nitrosylation of cysteine of 8000 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.
Upon receipt, the kit should be stored at -20°C. Please use within 6 months from the date of shipment.
