Analysis Note

UV assay: One unit will cause the oxidation of 1.0 µmole of NADPH at 25 °C at pH 7.5.Colorimetric assay: One unit will cause the reduction of 1.0 µmole of DTNB to TNB at 25 °C at pH 7.5.

Application

Glutathione Reductase Assay Kit has been used to measure the activity of glutathione reductase as a part of oxidative stress assessment and also to study the effects of antifouling biocides on it.

Biochem/physiol Actions

This kit provides reagents for a spectrophotometric assay for measuring the activity of glutathione reductase either by following the decrease in A₃₄₀ caused by the oxidation of NADPH or the increase in A₄₁₂ caused by the reduction of dithiobis(2-nitrobenzoic acid).

Glutathione reductase (EC 1.6.4.2) (GR) is a ubiquitous enzyme that catalyzes the reduction of oxidized glutathione (GSSG) to glutathione (GSH). Glutathione reductase is essential for the glutathione redox cycle that maintains adequate levels of reduced cellular GSH, which serves as an antioxidant reacting with free radicals and organic peroxides. Glutathione is also a substrate for the glutathione peroxidases and glutathione S-transferases in the detoxification of organic peroxides and the metabolism of xenobiotics. This kit contains reagents for the spectrophotometric determination of glutathione reductase activity either by following the decrease in absorbance caused by the oxidation of NADPH at 340 nm (UV assay) or the increase in absorption caused by the reduction of dithiobis(2-nitrobenzoic acid) (DTNB) at 412 nm (colorimetric assay). This kit provides reagents for a spectrophotometric assay for measuring the activity of glutathione reductase either by following the decrease in A₃₄₀ caused by the oxidation of NADPH or the increase in A₄₁₂ caused by the reduction of dithiobis(2-nitrobenzoic acid).

General description

Glutathione reductase (GR) is a ubiquitous enzyme that catalyzes the reduction of oxidized glutathione (GSSG) to glutathione (GSH). Glutathione reductase is essential for the glutathione redox cycle that maintains adequate levels of reduced cellular GSH, which serves as an antioxidant reacting with free radicals and organic peroxides. Glutathione is also a substrate for the glutathione peroxidases and glutathione S-transferases in the detoxification of organic peroxides and the metabolism of xenobiotics. 

Principle

This kit contains reagents for the spectrophotometric determination of glutathione reductase activity either by following the decrease in absorbance caused by the oxidation of NADPH at 340 nm (UV assay) or the increase in absorption caused by the reduction of dithiobis(2-nitrobenzoic acid) (DTNB) at 412 nm (colorimetric assay). This kit provides reagents for a spectrophotometric assay for measuring the activity of glutathione reductase either by following the decrease in A340 caused by the oxidation of NADPH or the increase in A412 caused by the reduction of dithiobis(2-nitrobenzoic acid).

Suitability

Suitable for the measurement of glutathione reductase activity in biological samples