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Specifications
- Assay Type Activity Assay
- Sample Types Serum, Plasma (EDTA and Heparin), Cell Lysates, RBCs
- Sensitivity 9 µU/mL
- Species Human
- Assay Duration 20 Minutes
- Samples/Plate 41 in Duplicate
- Readout Fluorescent, 510 nm emission / 370-410 nm excitation
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Standard Curve
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Description
Assay Principle:Â
The Glutathione Reductase (GR) Fluorescent Activity Kit quantitatively measure GR activity in serum, plasma (EDTA and Heparin), RBCs, and cell lysates. The Glutathione Reductase (GR) Fluorescent Activity Kit is an Activity Assay with a run time of 20 minutes. Please read the complete kit insert for more information before performing this assay.
Use our provided Glutathione Reductase Standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a black microtiter plate. Add ThioStar® Detection Reagent to each well tapping the plate to ensure sufficient mixing of reagents. Then incubate the mixture covered at room temperature for 5 minutes and record the fluorescent signal at 510nm to attain the background thiol content in the samples. Then add NADPH and Oxidized Glutathione (GSSG), tap the plate to mix, and incubate for another 15 minutes. The GR within the standard or sample will convert the oxidized glutathione, GSSG, into free GSH, which then reacts with the ThioStar® to yield the signal related to GR activity.
After the 15-minute incubation, use a plate reader to detect and record the generated fluorescent signal at 510nm. Use the intensity and the standard curve to calculate the GR activity in the samples.
Background:
Glutathione reductase (GR) is a ubiquitous 100-120 kDa dimeric flavoprotein encoded by the GSR gene. GR plays an indirect role in preventing oxidative damage within the cell. It catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH). GSH, in conjunction with glutathione peroxidase (GP), is the acting reductant responsible for minimizing harmful hydrogen peroxide. GSH dysregulation is correlated with diseases such as cancer, cystic fibrosis, and neurodegenerative diseases.