S-Glutathionylated Protein Detection Kit

Glutathione (GSH) is a tripeptide (?-glutamylcysteinylglycine) widely distributed in both plants and animals.{1813,4672} GSH is involved in maintenance of protein sulfhydryl reduction status.{4669,4670} The concentration of GSH ranges from a few micromolar in plasma to several millimolar in tissues such as liver.{4671,4673} Mixed protein glutathionyl disulfides are a post translational protein modification of growing interest.{11632,17080,15065} Protein S-glutathionylation (PSSG) may modify the activity of a large number of cell proteins, including metabolic, structural, cytoskeletal, and signaling proteins.{16795} PSSG detection methods can employ GSH adduct antibodies, GSH derivatives, and differential labeling systems based on the “Biotin-Switch” method.{15065,13441,12802} Cayman’s S-Glutathionylated Protein Detection Assay Kit provides a convenient method for the direct visualization of S-glutathionylated proteins in whole (permeabilized) cells by flow cytometry and microscopy as well as avidin overlay analysis.{15066} This cell-based assay starts with the modification of protein free-thiols groups followed by enzymatic cleavage of any protein-S-glutathione (PSSG) adducts present in the sample. Biotinylation of the newly-formed protein free-thiols provides the basis for visualization using streptavidin-based colorimetric or fluorescence detection. Reagents are provided to test three sets of 10 samples (most convenient) or up to thirty samples total at once if desired.