General description

Glutathione (GSH), a thiol-containing tripeptide (γ-glutamyl-cysteinyl-glycine), is a key antioxidant in many species. It has been highly implicated in the detoxification/elimination of xenobiotics (naturally occurring harmful compounds such as free radicals, hydroperoxides etc.) and in the maintenance of the oxidation state of protein sulfhydryl groups. In addition, GSH plays a pivotal role in the pathogenesis of numerous human diseases including cancer and cardio-vascular diseases. Glutathione is present in cells in both reduced (GSH) and oxidized (GSSG) forms- GSH being, the predominant species under normal physiological conditions inside cells. Thus, pathologic conditions causing oxidative stress would result in increased levels of GSSG. Therefore, the measurement of intracellular GSH appears to be a sensitive indicator of the overall cell health, and its ability of resisting toxic challenges.

Principle

The Reduced Glutathione (GSH) Assay Kit is based on an enzymatic cycling method in the presence of GSH and a chromophore. The reduction of the chromophore produces a stable product, which can be followed kinetically at A450 nm. Therefore, its absorbance is directly proportional to the amount of GSH in the sample. The kit is highly specific and sensitive because GSSG does not interfere with the assay. The kit includes 5-Sulfosalicylic acid (SSA) in order to protect samples’; GSH endogenous content. (SSA acts as a deproteinizing and antioxidant agent. The assay is simple, reproducible and can specifically detect as low as 50 pmol/well of reduced form of Glutathione (GSH) in a 100 µL reaction.

Suitability

The kit is suitable for the measurement of reduced glutathione in various biological samples/preparations including tissue homogenates (liver, etc.) and cell lysates (Hep G2, Jurkat, etc.).