Description
Key facts
Detection method:Colorimetric,
Sample types:Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Platelets, Other biological fluids,
Assay type:Enzyme activity,
Sensitivity:= 0.5 mU/mL,
Assay time:40m,
Assay Platform:Microplate reader
Product details:
How the assay works
Glutathione peroxidase assay kit measures the activity of glutathione peroxidase by measuring the decrease in absorbance of NADPH. The assay is based on the oxidation of reduced glutathione to oxidized glutathione.
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 the GSSG to produce GSH, and in the same reaction consumes NADPH. The decrease of NADPH (measured at OD=340 nm) is proportional to GPx activity.
Glutathione peroxidase assay protocol summary:
- Add samples and standards to wells
- Add reaction mix and incubate for 15 min at room temp to deplete all GSSG in sample
- Add cumene hydroperoxide
- Analyze with microplate reader immediately, and after at least 5 min
How other researchers are using
Glutathione Peroxidase Assay Kit (Colorimetric) has been used in a variety of sample type including:
Mouse C2C12 myoblasts
Mice heart samples
Rats gonads homogenates
References:
1-Martin S et al. 2023
2-Li H et al. 2023
3-Naguib G et al. 2023
Related and recommended products
For fluorometric detection, we recommend Glutathione Peroxidase Activity Assay Kit (Fluorometric)
ab219926
This product is manufactured by BioVision, an Abcam company and was previously called K762 Glutathione Peroxidase Activity Colorimetric Assay Kit. K762-100 is the same size as the 100 test size of ab102530.
The Glutathione Peroxidase (GPx, EC 1.11.1.9) family of enzymes plays and important role in the protection of 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.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Glutathione Peroxidase (GPx) often referred to as GSH peroxidase is an essential enzyme in the cellular defense system against oxidative damage. It exists as a group of enzymes including the widely studied isoform GPx4. GPx has a molecular weight of approximately 20-23 kDa varying slightly depending on the isoform. This enzyme is localized in many tissues across different organs including the liver kidney and muscles. It catalyzes the reduction of hydrogen peroxide and organic hydroperoxides using glutathione (GSH) as a substrate preventing the formation of free radicals.
Biological function summary
The enzyme plays a significant role in the detoxification of reactive oxygen species. GPx does not usually form a complex with other proteins; instead it acts individually within cells to maintain the balance of oxidative and reductive forces. By converting harmful peroxides into non-toxic alcohols and water it protects cell membranes and other cellular components from oxidative damage. GPx activity is often measured in laboratories using assays to determine its efficiency in catalyzing these reactions.
Pathways
GPx engages in critical cellular antioxidant defense mechanisms. It is an integral part of the glutathione redox cycle where it partners with glutathione reductase another enzyme important for regenerating reduced glutathione. Additionally GPx links to the pentose phosphate pathway providing reduced NADPH necessary for maintaining glutathione in its reduced form. These pathways protect cells from oxidative stress and keep redox homeostasis in check.
Alterations in GPx activity correlate with various health conditions. Researchers link decreased GPx activity to cardiovascular diseases and neurodegenerative disorders highlighting its importance in maintaining cellular health. In cardiovascular diseases the imbalance of oxidative stress pathways involves proteins like superoxide dismutase in addition to GPx worsening the effects of oxidative damage. In neurodegenerative disorders abnormal protein aggregations disturb GPx activities emphasizing the role of antioxidants in disease prevention and management.
