U-Blot® Human Protein S-Acylation Array G1

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.