U-Blot® Human Protein S-Nitrosylation Array G1

Protein S‑Nitrosylation plays an unusually prominent role in cell signaling, development and growth. The G‑Series Human Protein S‑Nitrosylation Antibody Array 1 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. This multiplex glass slide array is specifically designed for simultaneous identification of the relative levels of S‑Nitrosylation of cysteine of 507 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. The 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.