Glutathione Assay Kit, Fluorimetric

Application

Glutathione Assay Kit, Fluorimetric has been used in intracellular thiol assays and in the measurement of hepatic glutathione in rats.

The Glutathione Assay Kit, Fluorimetric includes all the reagents required for fast and easy measurement of reduced glutathione in cell or tissue extracts or directly in growing cells. Staurosporine is supplied with the kit to enable induction of apoptosis in cells. Since the amount of reduced glutathione in staurosporine-induced cells is significantly lower than in non-induced cells, this can serve as a control system.

Biochem/physiol Actions

Glutathione (GSH) acts as a cofactor for several antioxidant enzymes. It helps in the regeneration of vitamins C and E. It regulates cellular proliferation and apoptosis. GSH is important for mitochondrial functioning and the maintenance of mitochondrial DNA (mtDNA). Mutations in this gene is associated with Alzheimer′s, Parkinson′s and Huntington′s diseases.

Reduced glutathione (GSH), a tripeptide (γ-glutamyl-cysteinylglycine), is the major free thiol in most living cells and is involved in many biological processes such as detoxification of xenobiotics, removal of hydroperoxides, and maintenance of the oxidation state of protein sulfhydryls. It is the key antioxidant in animal tissues. Glutathione is present inside cells primarily in the reduced form (90-95% of the total glutathione). The remainder is present in the oxidized form (glutathione disulfide, GSSG). Intracellular GSH status appears to be a sensitive indicator of the overall health of a cell and its ability to resist toxic challenge. High levels of GSH in the cell may indicate pathological changes.

Analysis Note

The kit assay utilizes a thiol probe (monochlorobimane), which can freely pass through the plasma membrane. The free, unbound probe shows very little fluorescence, but when bound to reduced glutathione in a reaction that is catalyzed by glutathione S-transferase (GST), it forms a strongly fluorescent adduct.