Assay Principle:

The DetectX® Glutathione Reductase (GR) Fluorescent Activity Kit is designed for the quantitative measurement of GR activity in serum, plasma (EDTA and Heparin), RBCs, and cell lysates. The kit offers a rapid assay process with a run time of 20 minutes. For optimal results, it is important to follow the detailed instructions provided in the kit insert. The kit includes a Glutathione Reductase Standard to create a precise standard curve.

Protocol Summary:

• Introduce standards or diluted samples into the black microtiter plate.
• Add ThioStar® Detection Reagent to each well, ensuring thorough mixing.
• Incubate the mixture at room temperature for 5 minutes, then measure the background thiol content at 510nm.
• Add NADPH and Oxidized Glutathione (GSSG), mix well, and incubate for an additional 15 minutes. GR in the sample will convert GSSG to GSH, which then reacts with ThioStar® to indicate GR activity.
• After incubation, use a plate reader to detect and measure the fluorescent signal at 510nm. Calculate the GR activity using the standard curve.

Background:

Glutathione reductase (GR), a dimeric flavoprotein, is encoded by the GSR gene and is found in various tissues. It plays a key role in cellular defense against oxidative damage by catalyzing the reduction of oxidized glutathione (GSSG) to its reduced form (GSH). GSH, with glutathione peroxidase (GP), acts as a reductant, mitigating the effects of harmful hydrogen peroxide within cells.

Clinical researchers hve linked a range of diseases, including cancer, cystic fibrosis, and neurodegenerative disorders to dysregulation of GSH. Therefore, understanding GR activity is essential in research focusing on oxidative stress and related diseases. Accurate measurement of GR activity can provide insights into the cellular redox balance and contribute to the development of therapeutic strategies for conditions associated with oxidative stress.

The DetectX® Glutathione Reductase (GR) Fluorescent Activity Kit, with its efficient and sensitive detection capabilities, is an important resource for researchers studying the mechanisms of oxidative stress and exploring potential interventions for diseases where GSH and GR play major roles.