Product name: Glutathione Peroxidase Assay Kit (Colorimetric)
Detection method: Colorimetric
Example type: Cell culture supernatant, urine, serum, plasma, platelets, other biological fluids, tissue extracts
Test type: Enzymatic activity
Sensitivity: 0.5 mU/ml
Test time: 0h 40m
Reacts with: Mammals, Other species
The ab102530 Glutathione Peroxidase Assay Kit (Colorimetric) can be used to quantify the activity of all glutathione-dependent peroxidases in plasma, erythrocyte lysates, tissue homogenates, and cell lysates. In the glutathione peroxidase assay protocol, glutathione peroxidase (GPx) oxidizes GSH to produce GSSG as part of the reaction in which it reduces cumene hydroperoxide. Glutathione reductase (GR) then reduces GSSG to produce GSH, and in the same reaction consumes NADPH. The decrease in NADPH (measured at OD = 340 nm) is proportional to the activity of GPx. The assay has a detection sensitivity of ~ 0.5 mU/ml GPx in samples.
Summary of the glutathione peroxidase assay protocol:
– add samples and standards to wells
– add reaction mix and incubate for 15 minutes at room temperature to deplete all GSSGs in the sample
– add cumene hydroperoxide
– analyze with a microplate reader immediately and after at least 5 min
The glutathione peroxidase family of enzymes (GPx, EC 18.104.22.168) plays an important role in protecting organisms from oxidative damage. GPx converts reduced glutathione (GSH) to oxidized glutathione (GSSG) while reducing lipid hydroperoxides to their corresponding alcohols or free hydrogen peroxide to water.
Glutathione peroxidase (GPx, EC 22.214.171.124) is a family of enzymes with peroxidase activity and plays an important role in protecting organisms from oxidative damage. Converts reduced glutathione (GSH) to oxidized glutathione (GSSG), to reduce lipid hydroperoxides to their corresponding alcohols, or to reduce free hydrogen peroxide in water. Several isozymes have been found in different cell locations and with different substrate specificity. Low levels of GPx have been correlated with free radical-related disorders.