CheKine™ Micro Glutathione Reductases (GR) Activity Assay Kit

GR is a flavin protein redox enzyme widely present in eukaryotes and prokaryotes. GR catalyzes the reduction of GSSG to GSH. It is one of the key enzymes in the glutathione redox cycle (generally GR is replaced by TrxR in insects) . GR catalyzes the reduction of GSSG by NADPH to GSH, which helps maintain the GSH / GSSG ratio in the body. GR plays a key role in scavenging reactive oxygen species in response to oxidative stress, and GR is also involved in the ascorbate-glutathione cycle pathway.

The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.

Storage at -20°C and Keep from light immediately upon receipt. Kit has a storage time of 12 months from receipt. Refer to list of materials supplied for storage conditions of individual components.

GR can catalyze the reduction of NADPH to GSSG to regenerate GSH, and NADPH dehydrogenates to produce NADP +; NADPH has a characteristic absorption peak at 340 nm, while NADP + has no absorption peak at this wavelength; NADPH dehydrogenation rate is determined by measuring the decrease rate of absorbance at 340 nm to calculate GR activity.

• 96 Well Clear Flat Bottom UV-Transparent Microplate is needed. • The whole processes need to be carried out on ice, and the enzyme activity should be determined on the same day, to avoid repeated freeze-thaw of the homogenate solution. • In the detection of GR activity in cells, the cell number must be between 3-5 x 106, and the extraction of GR in cells can be followed by Assay Buffer grinding or ultrasonic treatment, cells cannot be treated by lysate. • Before measurement, one or two samples should be used as a pre-experiment to ensure that the change in absorbance within 180 s is linear. • Generally, mammalian tissue must be diluted 2 to 5 times with Assay Buffer. • The measuring process should be operated quickly and only 1 ~ 2 blank well should be needed. • The reaction temperature has influence on the result. Keep the temperature at 25°C (for general species) or 37°C (for mammals) .