Glutathione Reductase Assay Kit

Trevigen’s Glutathione Reductase Assay Kit is a higher volume assay designed to measure cellular levels of Glutathione Reductase, an enzyme which catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH) (figure1). GSH is an important intracellular antioxidant that reacts with oxygen free radicals and organic peroxides and acts as a substrate for other detoxification enzymes such as glutathione peroxidase and glutathione S-transferase. The activity of glutathione reductase is an important measure of the antioxidant status of the cell. Glutathione reductase requires NADPH for its activity, resulting in the reduction of GSSG to GSH and the corresponding oxidation of NADPH to NADP+.  

Trevigen’s Glutathione Reductase Assay takes less than 6 minutes to perform and is based on the measurement of the loss in absorbance at 340 nm due to the conversion of NADPH to NADP+. The kit contains everything that you need to perform the assay on 100 samples and its corresponding blanks, each in duplicate, as well as 10 standard curves with each point performed in duplicate. In total, each kit can generate data on over 500 data points. The NADPH is provided lyophilized in 10 vials, each containing sufficient NADPH for generating one standard curve and for determining glutathione reductase levels in 10 experimental samples.

HT Glutathione Assay Kit

Trevigen’s Glutathione Assay utilizes a carefully optimized enzymatic recycling method, which employs Glutathione Reductase, for the quantification of glutathione. The sulfhydryl group of glutathione reacts with DTNB (5,5’-dithiobis-2-nitrobenzoic acid, Ellman’s reagent) and produces a yellow-colored 5-thio-2-nitrobenzoic acid (TNB). The mixed disulfide, GSTNB (between GSH and TNB) that is concomitantly produced, is reduced by Glutathione Reductase to recycle the GSH and produce more TNB. The rate of TNB production is directly proportional to the concentration of GSH in the sample. Thus, measurement of the absorbance of TNB at 405 or 412 nm provides an accurate estimation of the amount of glutathione in the sample.  

HT Glutathione Peroxidase Assay Kit

Glutathione peroxidase (GP) is found in mammalian cells and helps to prevent lipid peroxidation of cell membranes by consuming free peroxide in the cell. Thus, Glutathione Peroxidase plays a critical role in protecting the cell from free radical damage. Tetrameric Glutathione Peroxidase catalyzes the reduction of H2O2 to water, and organic peroxides to the corresponding stable alcohols, by using glutathione as a source of reducing equivalents. It requires selenium as a cofactor and contains a selenocysteine amino acid residue in the active site of each monomer that participates in the actual mechanism of the enzyme. Glutathione Reductase (GR), provided with each kit, then reduces the oxidized glutathione to complete the cycle.  The oxidation of NADPH to NADP+ is accompanied by a decrease in absorbance at 340 nm. The rate of decrease in the absorbance at 340 nm is directly proportional to the Glutathione Peroxidase activity in the sample. Trevigen’s Glutathione Peroxidase Assay Kit can be used to measure glutathione-dependent peroxidases in plasma, erythrocyte lysates, tissue homogenates, and cell lysates.

HT Glutathione Reductase Assay Kit

This high throughput glutathione reductase assay kit is designed to measure cellular levels of Glutathione Reductase, an enzyme which catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH) (Figure 1). GSH is an important intracellular antioxidant that reacts with oxygen free radicals and organic peroxides and acts as a substrate for other detoxification enzymes such as glutathione peroxidase and glutathione S-transferase. The activity of glutathione reductase is an important measure of the antioxidant status of the cell. Glutathione reductase requires NADPH for its activity, resulting in the reduction of GSSG to GSH and the corresponding oxidation of NADPH to NADP+.

Trevigen's Glutathione Reductase Assay Kit is a spectrophotometric assay in which the oxidation of NADPH to NADP+ is monitored by the decrease in absorbance at 340 nm. This rate of decrease in absorbance at 340 nm is directly proportional to the glutathione reductase activity in the sample because the enzyme is present at rate limiting concentrations. This kit provides the user with all the reagents and plates to easily and rapidly assay for glutathione reductase in cell and tissue extracts.