The NWLSS™ Glutathione Assay kit is a kinetic, rate based assay for quantification of total GSH (reduced and oxidized) in a variety of samples including blood, plasma, tissues and cultured cells. Suitable for measurement of oxidized GSH (GSSG) after scavenging of reduced GSH.
Recommended for researchers wishing to assay GSH or GSSG in tissue homogenates, cell lysates & other biological fluid where GSH is present.
This assay is compatible with samples from all species.
Glutathione (GSH, g-glutamylcysteinylglycine), the primary non-protein sulfhydryl in aerobic organisms is synthesized in most cells. The ubiquitous tripeptide is formed by the combination of glutamic acid and cysteine, catalyzed by g-glutamylcysteinyl synthetase. Glycine is then added by glutathione synthetase to form GSH.
In addition to donating an electron during the reduction of hydroperoxides to the respective alcohols (or water in the case of hydrogen peroxide), GSH functions as a co-substrate in the metabolism of xenobiotics catalyzed by glutathione S-transferases. It is also a co-factor for several metabolic enzymes is involved in intracellular transport, functions as an antioxidant and radioprotectant and facilitates protein folding and degradation.
The NWLSS™ Glutathione Assay is a modification of the method first described by Tietze. The general thiol reagent, 5-5'-dithiobis[2-nitrobenzoic acid] (DTNB, Ellman's Reagent) reacts with GSH to form the 412 nm chromophore, 5-thionitrobenzoic acid (TNB) and GS-TNB. The GS-TNB is subsequently reduced by glutathione reductase and β-nicotinamide adenine dinucleotide phosphate (NADPH), releasing a second TMB molecule and recycling the GSH; thus amplifying the response. Any GSSG initially present in the reaction mixture or formed from the mixed disulfide reaction of GSH with GS-TNB is rapidly reduced to GSH and detectable in the assay.
View Assay Protocol
96 tests presented as 1 X 96 well microtiter plate assay. Cuvette format for spectrophotometers also available.
Quantitative measurement of total GSH (reduced plus oxidized form)
Whole blood, plasma, tissues and cell lysates.
Total Glutathione (GSH plus GSSG)
0.1 µM using standard method (for whole blood & some tissue types). 0.005 µM using modified high sensitivity method (for samples with lower GSH levels such as plasma or for detection of GSSG after reduced GSH has been scavenged). Contact NWLSS for details.
|Storage and Stability:||
12 months from date of manufacture if stored at 2-8C
GSH (GSSG) Calibrator