Recombinant Anti-NQO1 antibody [EPR3309] (ab80588)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3309] to NQO1
- Suitable for: WB, IP, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Rat, Human
Overview
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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: Rat, Human
Predicted to work with: Mouse
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Immunogen
Synthetic peptide within Human NQO1 (N terminal). The exact sequence is proprietary.
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Positive control
- WB: SH-SY5Y, MCF7, HeLa and A549 cell lysates, rat brain tissue lysate. ICC/IF: MCF-7 cells. IP: HeLa cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3309 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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.
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| Flow Cyt | Use at an assay dependent concentration. | |
| ICC/IF | 1/50 - 1/80.
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Target
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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
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Database links
- Entrez Gene: 1728 Human
- Entrez Gene: 18104 Mouse
- Entrez Gene: 24314 Rat
- Omim: 125860 Human
- SwissProt: P15559 Human
- SwissProt: Q64669 Mouse
- SwissProt: P05982 Rat
- Unigene: 406515 Human
see all -
Alternative names
- Azoreductase antibody
- Cytochrome b 5 reductase antibody
- DHQU antibody
see all
Images
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![Western blot - Anti-NQO1 antibody [EPR3309] (ab80588)](https://www.abcam.com/ps/products/80/ab80588/Images/ab80588-297796-ab80588KOWBJRIZBranford.jpg)
Western blot - Anti-NQO1 antibody [EPR3309] (ab80588)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 kDaLanes 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.
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![Western blot - Anti-NQO1 antibody [EPR3309] (ab80588)](https://www.abcam.com/ps/products/80/ab80588/Images/ab80588-239097-ab80588pwb.jpg)
Western blot - Anti-NQO1 antibody [EPR3309] (ab80588)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 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-NQO1 antibody [EPR3309] (ab80588)](https://www.abcam.com/ps/products/80/ab80588/Images/ab80588-239101-ab80588piccif.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-NQO1 antibody [EPR3309] (ab80588)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).
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![Flow Cytometry - Anti-NQO1 antibody [EPR3309] (ab80588)](https://www.abcam.com/ps/products/80/ab80588/Images/ab80588-269571-ab80588FC1.jpg)
Flow Cytometry - Anti-NQO1 antibody [EPR3309] (ab80588)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)
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![Immunoprecipitation - Anti-NQO1 antibody [EPR3309] (ab80588)](https://www.abcam.com/ps/products/80/ab80588/Images/ab80588-239099-ab80588pip.jpg)
Immunoprecipitation - Anti-NQO1 antibody [EPR3309] (ab80588)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.
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![Western blot - Anti-NQO1 antibody [EPR3309] (ab80588)](https://www.abcam.com/ps/products/80/ab80588/Images/NQO1-Primary-antibodies-ab80588-1.jpg)
Western blot - Anti-NQO1 antibody [EPR3309] (ab80588)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
Protocols
References (32)
ab80588 has been referenced in 32 publications.
- Yao X et al. PTEN inhibitor VO-OHpic attenuates GC-associated endothelial progenitor cell dysfunction and osteonecrosis of the femoral head via activating Nrf2 signaling and inhibiting mitochondrial apoptosis pathway. Stem Cell Res Ther 11:140 (2020). PubMed: 32228695
- Zheng Y et al. Desflurane Preconditioning Protects Against Renal Ischemia-Reperfusion Injury and Inhibits Inflammation and Oxidative Stress in Rats Through Regulating the Nrf2-Keap1-ARE Signaling Pathway. Drug Des Devel Ther 14:1351-1362 (2020). PubMed: 32308368
- Yu H et al. Size and temporal-dependent efficacy of oltipraz-loaded PLGA nanoparticles for treatment of acute kidney injury and fibrosis. Biomaterials 219:119368 (2019). PubMed: 31349200
- Takashima M et al. Neuroprotective effects of Brazilian green propolis on oxytosis/ferroptosis in mouse hippocampal HT22 cells. Food Chem Toxicol 132:110669 (2019). PubMed: 31299294
- Zhang J et al. p62 functions as an oncogene in colorectal cancer through inhibiting apoptosis and promoting cell proliferation by interacting with the vitamin D receptor. Cell Prolif N/A:e12585 (2019). PubMed: 30793399
