Key features and details
- HRP Mouse monoclonal [A180] to NQO1
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
- Conjugation: HRP
- Isotype: IgG1
Product nameHRP Anti-NQO1 antibody [A180]
See all NQO1 primary antibodies
DescriptionHRP Mouse monoclonal [A180] to NQO1
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
Recombinant full length protein corresponding to Human NQO1.
- WB: Human kidney tissue lysate. IHC-P: human breast adenocarcinoma tissue sections.
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We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. Store In the Dark.
Storage bufferpH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: 30% Glycerol, 1% BSA, PBS
Concentration information loading...
ELISA pair antibody
Our Abpromise guarantee covers the use of ab196629 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||1/5000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).|
FunctionThe enzyme apparently serves as a quinone reductase in connection with conjugation reactions of hydroquinons involved in detoxification pathways as well as in biosynthetic processes such as the vitamin K-dependent gamma-carboxylation of glutamate residues in prothrombin synthesis.
Sequence similaritiesBelongs to the NAD(P)H dehydrogenase (quinone) family.
- Information by UniProt
- Azoreductase antibody
- Cytochrome b 5 reductase antibody
- DHQU antibody
All lanes : HRP Anti-NQO1 antibody [A180] (ab196629) at 1/5000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : NQO1 knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 30 kDa
Observed band size: 31 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutes
ab196629 was shown to recognize NQO1 in wild-type HAP1 cells as signal was lost at the expected MW in NQO1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NQO1 knockout samples were subjected to SDS-PAGE. Ab196629 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.
IHC image of NQO1 staining in a section of formalin-fixed paraffin-embedded human breast adenocarcinoma*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab196629 at 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
HRP Anti-NQO1 antibody [A180] (ab196629) at 1/5000 dilution + Human kidney tissue lysate - total protein (ab30203) at 20 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 30 kDa
Exposure time: 20 minutes
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab196629 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
ab196629 has not yet been referenced specifically in any publications.