Overview

  • Product name
    Anti-NQO1 antibody [EPR3309]
    See all NQO1 primary antibodies
  • Description
    Rabbit monoclonal [EPR3309] to NQO1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human NQO1 (N terminal). The exact sequence is proprietary.

  • Positive control
    • WB: SH-SY5Y, MCF7, HeLa and A549 cell lysates, rat brain tissue lysate. ICC/IF: MCF-7 cells. IP: HeLa cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab80588 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/10000 - 1/150000. Detects a band of approximately 31 kDa (predicted molecular weight: 31 kDa).
IP 1/30 - 1/50.

 

Flow Cyt Use at an assay dependent concentration.
ICC/IF 1/50 - 1/80.

 

Target

  • Function
    The 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 similarities
    Belongs to the NAD(P)H dehydrogenase (quinone) family.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • Azoreductase antibody
    • Cytochrome b 5 reductase antibody
    • DHQU antibody
    • DIA 4 antibody
    • DIA4 antibody
    • Diaphorase (NADH/NADPH) (cytochrome b 5 reductase) antibody
    • Diaphorase (NADH/NADPH) antibody
    • Diaphorase 4 antibody
    • Dioxin inducible 1 antibody
    • DT diaphorase antibody
    • DT-diaphorase antibody
    • DTD antibody
    • Menadione reductase antibody
    • NAD(P)H dehydrogenase [quinone] 1 antibody
    • NAD(P)H dehydrogenase quinone 1 antibody
    • NAD(P)H menadione oxidoreductase 1 dioxin inducible antibody
    • NAD(P)H quinone dehydrogenase 1 antibody
    • NAD(P)H: menadione oxidoreductase 1 dioxin inducible 1 antibody
    • NAD(P)H:menadione oxidoreductase 1 antibody
    • NAD(P)H:Quinone acceptor oxidoreductase type 1 antibody
    • NAD(P)H:quinone oxidoreductase 1 antibody
    • NAD(P)H:quinone oxireductase antibody
    • NMOR 1 antibody
    • NMOR I antibody
    • NMOR1 antibody
    • NMORI antibody
    • NQO 1 antibody
    • NQO1 antibody
    • NQO1_HUMAN antibody
    • Phylloquinone reductase antibody
    • QR 1 antibody
    • QR1 antibody
    • Quinone reductase 1 antibody
    see all

Images

  • All lanes : Anti-NQO1 antibody [EPR3309] (ab80588) at 1 µg/ml

    Lane 1 : Wild-type HAP1 whole cell lysate at 40 µg
    Lane 2 : NQO1 knockout HAP1 whole cell lysate at 40 µg
    Lane 3 : HepG2 whole cell lysate at 20 µg
    Lane 4 : MEF1 whole cell lysate at 20 µg
    Lane 5 : Human Kidney tissue lysate at 20 µg
    Lane 6 : Mouse Kidney tissue lysate at 20 µg

    Predicted band size: 31 kDa



    Lanes 1 - 6: Merged signal (red and green). Green - ab80588 observed at 31 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab80588 was shown to recognize NQO1 in wild-type HAP1 cells as signal was lost at the expected MW in NQO1 knockout cells. Wild-type and NQO1 knockout samples were subjected to SDS-PAGE. Ab80588 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-NQO1 antibody [EPR3309] (ab80588) at 1/25000 dilution (purified)

    Lane 1 : SH-SY5Y cell lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 31 kDa
    Observed band size: 31 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunocytochemsitry/Immunofluorescence analysis of MCF-7 cells labelling NQO1 (green) with purified ab80588 at 1/80. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

  • ab80588 staining NQO1 in the human cell line MCF-7(human breast carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • ab80588 (purified) at 1/50 immunoprecipitating NQO1 in HeLa cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-NQO1 antibody [EPR3309] (ab80588) at 1/50000 dilution (unpurified)

    Lane 1 : MCF7
    cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : A549
    cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 31 kDa
    Observed band size: 31 kDa

References

This product has been referenced in:
  • Wible RS  et al. NRF2 regulates core and stabilizing circadian clock loops, coupling redox and timekeeping inMus musculus. Elife 7:N/A (2018). Read more (PubMed: 29481323) »
  • Cheng X  et al. Enhanced glycometabolism as a mechanism of NQO1 potentiated growth of NSCLC revealed by metabolomic profiling. Biochem Biophys Res Commun 496:31-36 (2018). Read more (PubMed: 29291405) »

See all 12 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (MDA-MB-468 cells)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH6
Permeabilization
No
Specification
MDA-MB-468 cells
Blocking step
No blocking step used for 1 hour(s) and 0 minute(s) · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted May 31 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (B16 tumor lysate)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
B16 tumor lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Apr 09 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MDA-MB-468 cell lysate)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
MDA-MB-468 cell lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Apr 09 2018

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up