Glutathione reductase (GR) plays an indirect but essential role in the prevention of oxidative damage within the cell by helping to maintain appropriate levels of intracellular glutathione (GSH). GSH is the acting reductant responsible for minimizing harmful hydrogen peroxide cellular levels. The regeneration of GSH is catalyzed by GR. GR is an ubiquitous 100-120 kDa dimeric flavoprotein that catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione, using NADPH as the hydrogen donor. Molecules such as NADPH act as hydride donors in a variety of enzymatic processes. NADPH has been suggested to also act as an indirectly operating antioxidant, given its role in the re-reduction of GSSG to GSH and thus maintaining the antioxidative power of glutathione.
The Glutathione Reductase (GR) Activity kit is designed to quantitatively measure GR activity in a variety of samples. A GR standard is provided to generate a standard curve for the assay. The kit utilizes a proprietary non-fluorescent molecule, to covalently bind to the free thiol group on GSH, the product of the GR-catalyzed GSSG reduction, to yield a highly fluorescent product. After mixing the sample or standard with and incubating at room temperature, the fluorescent product is read at 510 nm in a fluorescent plate reader with excitation at 390 nm.
Serum, plasma, erythrocytes and cell lysate samples have been validated with this assay.
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