NQO1 Activity Assay Kit (ab184867)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 30 min
- Sample type: Adherent cells, Cell culture extracts, Cell Lysate, Suspension cells, Tissue Extracts, Tissue Homogenate, Tissue Lysate
Overview
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Product name
NQO1 Activity Assay Kit -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Overall 3 2.183% Inter-assay Sample n Mean SD CV% Overall 9 6.035% -
Sample type
Cell culture extracts, Adherent cells, Suspension cells, Tissue Extracts, Cell Lysate, Tissue Homogenate, Tissue Lysate -
Assay type
Enzyme activity -
Assay time
0h 30m -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human -
Product overview
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 -
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.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests 1000X Cofactor (lyophilized) 1 vial 1000X Inhibitor (lyophilized) 1 vial 100X Dye (lyophilized) 1 vial 20X Basic Buffer 1 x 3ml 2X Extraction Buffer 1 x 15ml 5000X Menadione 1 x 100µl 500X NADH (lyophilized) 1 vial 96-well Microplate 2 units -
Research areas
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Cellular localization
Cytoplasmic -
Alternative names
- Azoreductase
- DIA4
- DT-diaphorase
see all -
Database links
- Entrez Gene: 519632 Cow
- Entrez Gene: 1728 Human
- Entrez Gene: 18104 Mouse
- Entrez Gene: 24314 Rat
- SwissProt: P15559 Human
- SwissProt: Q64669 Mouse
- SwissProt: P05982 Rat
Images
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Raw data from various concentrations of HepG2 cell extracts.
An example is shown below where the rate/slope is calculated between these time points.
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NQO1 Activity in HepG2 extract.This raw data is expressed as rate (mOD/min) per microgram of cell extract added per well as shown above. The molar extinction coefficient factor for the WST1 dye is 25.9/mM/well.
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Raw data of NQO1 activity from 50 µg/mL of a series of normal samples.Figure 3a
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Comparison of NQO1 activity in different samples at 50 ug/mLFigure 3b
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Dicoumarol sensitive NQO1 activity/total activity in a series of normal cell lysates at 50 ug/mL.Figure 3c
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NQO1 activity assay components requirement test.Figure 4