Glutathione Assay Kit | GSH assay kit

Glutathione (GSH) Assay Kit

Cat. Num. AAA480438

• Synonyms: Glutathione; Glutathione (GSH) Assay Kit; Colorimetric Assay for Glutathione; GSH assay kit

• Preparation and Storage: 4 degree C

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Introduction: The Analyte
Glutathione (gamma-glutamylcysteinylglycine or GSH) is a naturally occuring tripeptide whose
nucleophilic and
reducing properties play a central role in metabolic pathways, as well as in the antioxidant system of most
aerobic
cells.1 GSH plays a critical role as a coenzyme with a variety of enzymes including, glutathione peroxidase,
glutathione S-transferase, and thiol transferase. GSH also plays major roles in drug metabolism, calcium
metabolism, the ?-glutamyl cycle, blood platelet, and membrane functions. In addition, GSH is crucial to a
variety of
life processes, including the detoxification of xenobiotics, maintenance of the -SH level of proteins, thiol-
disulfide
exchange, removal of hydroperoxides and free radicals, and amino acid transport across membranes.
Physiological
values of intracellular GSH generally range from 1 to 10 mM. Although many methods have been
described for the
assay of GSH, the reliable ones are labor intensive and not easy to use.2).
Principles of the Procedures
This method is based on a chemical reaction which proceeds in two steps. The first step leads to the
formation of
substitution products (thioethers) between a patented reagent, R1 (4-chloro-1-methyl-7-trifluromethyl-
quinolinium
methylsulfate), and all mercaptans (RSH) which are present in the sample:
The second step is a ?-elimination reaction which takes place under alkaline conditions. This reaction is
mediated by
reagent R2 (30% NaOH) which specifically transforms the substitution product (thioether) obtained with
GSH into a
chromophoric thione which has a maximal absorbance wavelength at 400 nm:
This method makes it possible to specifically assay glutathione with only one sampling and one
colorimetric
measurement. A modification of this method can be used to assay other mercaptans. This is based on the
measurement of substitution products, thioethers, which absorb light at 356 nm in the absence of reagent
R
2.
Because of its simplicity, this method is especially well adapted to the assay of glutathione in large series
of
biological samples. The main advantage of the method is the specificity for glutathione and it does not
require an
enzyme as a reagent.

References

1. Dolphin D., Poulson R. and Avramovic O., Eds. (1989), Glutathione: Chemical, Biochemical and Medical
Aspects, Vols A & B, J. Wiley and Sons.
2. Anderson M.E. (1989), Enzymatic and chemical methods for the determination of Glutathione; In:
Glutathione:
chemical, biochemical and medical aspects, Vol. A, Dolphin D., Poulson R. and Avramovic O. Eds., John
Wiley and Sons, pp.339-365.