- Detection method: Colorimetric method
Detection principle
Glutathione peroxidase (GSH-Px) can promote the reaction of hydrogen peroxide (H2O2) and reduced glutathione to produce H2O and oxidized glutathione (GSSG). The activity of glutathione peroxidase can be expressed by the rate of enzymatic reaction. The activity of glutathione can be calculated by measuring the consumption of reduced glutathione. Hydrogen peroxide (H2O2) and reduced glutathione can react without catalysis of GSH-Px, so the portion of GSH reduction by non-enzymatic reaction should be subtracted. GSH can react with dinitrobenzoic acid to produce 5-thio-dinitrobenzoic acid anion, which showed a stable yellow color. Measure the absorbance at 412nm, and calculate the amount of GSH.
Performance characteristics
Synonyms |
GSH-PX |
Sample type |
Serum,plasma,cells,cell culture supernatant,tissue |
Sensitivity |
12.65 U |
Detection range |
12.65-387 U |
Detection method |
Colorimetric method |
Assay type |
Enzyme Activity |
Assay time |
70 min |
Precision |
Average inter-assay CV: 9.300%Average intra-assay CV: 4.900% |
Other instruments required |
Micropipettor, Incubator, Vortex mixer, Centrifuge |
Other reagents required |
Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
Storage |
2-8℃ |
Valid period |
12 months |
N V Mohamad et al investigate the therapeutic effect of annatto tocotrienol for postmenopausal bone loss. Glutathione peroxidase (GSH-Px) activity of rat serum was determined using GSH-Px activity assay kit (E-BC-K096-S).
Dilution of sample
The optimal sampling volume are different for different species, also are different for different sample type. It is recommended to take 2~3 samples to do a pre-experiment, diluting a series of diluent and determine the dilution factor when the inhibition ratio is 20%~60% (the optimal inhibition ratio is the range of 45%~55%.) before formal experiment.
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).