Diminished cellular glutathione (GSH) level occurs at the early stage of mitochondrion-associated apoptosis pathway due to GSH efflux. GSH depletion further leads to cytochrome c release and caspase 3 induction (1,2). MCB Glutathione Detection Kit utilizes a thiol-reactive dye monochlorobimane (MCB), which is essentially nonfluorescent until it reacts with a thiol to form a blue fluorescent product (λAbs/λEm = 380/461 nm). By incubating cell lysate with MCB, the intensity of the fluorescent signal generated from the assay reflects the amount of GSH present in the cells. Fluorescence can easily be detected using a fluorometer or 96-well fluorometric plate reader with approximately 1 hour assay time.
Cell lysis buffer
GST positive control