Description
Key facts
Detection method:Colorimetric,
Sample types:Tissue Lysate, Cell culture extracts, Suspension cells, Tissue Extracts, Tissue Homogenate, Adherent cells, Cell Lysate,
Reacts with:Mouse, Rat, Cow, Human,
Assay type:Enzyme activity,
Assay time:30m,
Assay Platform:Microplate reader
Product details:
NQO1 Activity Assay kit (ab184867) is designed for the sensitive and accurate measurement of NQO1 (NAD(P)H dehydrogenase [quinone] 1) activity.
The enzyme activity is determined by following the reduction of Menadione with cofactor NADH and the simultaneous reduction of WST1 which leads to increased absorbance at 440 nm. Dicoumarol is used as an inhibitor of NQO1 as part of the assay.
NQO1 assay protocol summary:
- add samples to wells
- add reaction buffer + inhibitor and reaction buffer to separate sample wells
- analyze with a microplate reader for 5 min
Other Notes
NQO1 serves as a quinone reductase in connection with conjugation reactions of hydroquinones involved in detoxification pathways, NQO1 protects against quinone-induced damage by competing with potentially toxic one-electron pathways. It also functions in biosynthetic processes such as the vitamin K-dependent gamma-carboxylation of glutamate residues in prothrombin synthesis.
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.
NQO1 also known as NAD(P)H:quinone oxidoreductase 1 is a cytosolic enzyme involved in two-electron reduction processes. This protein plays a role in detoxification transforming quinones into less reactive and harmful hydroquinones. NQO1 has a molecular weight of approximately 31 kDa and is present in various tissues with high expression in the liver and lungs. Sometimes called DT-diaphorase it contributes to antioxidant protection within cells and assists in the stabilization of other proteins such as p53.
Biological function summary
NQO1 serves a protective function against oxidative stress by reducing quinones and preventing redox cycling that generates reactive oxygen species. Part of a critical network its function integrates with the cellular defense mechanism assisting in maintaining cellular homeostasis. NQO1 helps metabolize xenobiotics and is associated with phase II detoxification working alongside enzymes like glutathione S-transferases but is not part of a complex.
Pathways
NQO1 is an important component of the antioxidant defense pathway participating in the direct enzymatic reduction of quinones protecting cells from oxidative damage. It also interfaces with the KEAP1-NRF2 pathway where the NQO1 gene is regulated by the NRF2 transcription factor that upregulates its expression in response to oxidative stress. NQO1 interlinks with proteins such as p53 through pathways related to apoptotic regulation and cellular stress responses.
Mutations or altered expression of NQO1 have correlations with cancer development and progression notably in liver and lung cancers. This protein's stability influences cancer cell survival particularly under oxidative stress or chemotherapeutic treatments. NQO1 also exhibits links to Alzheimer's disease where its potential role in neuroprotection against oxidative damage draws investigation relating it to proteins like tau and amyloid-beta.
