Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab28947 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
Sandwich ELISA 1/500. Can be used as capture antibody in conjunction with ab34173 as detection antibody.
IP Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 30 kDa.
ICC/IF Use a concentration of 1 µg/ml.
ELISA Use at an assay dependent concentration. Can be paired for ELISA with Rabbit polyclonal to NQO1 (ab34173).
IHC-P Use at an assay dependent concentration.

Target

  • 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.
  • Cellular localizationCytoplasm.
  • 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: 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

Anti-NQO1 antibody [A180] images

  • IHC image of NQO1 staining in sections of formalin fixed paraffin embedded normal human pancreas* (left) and human pancreas adenocarcinoma* (right), performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28947, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    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

  • IHC image of NQO1 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28947, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    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.

  • Human breast cancer tissue stained with ab28947 NQO1 antibody.
  • Lane 1 : Marker
    Lane 2 : Anti-NQO1 antibody [A180] (ab28947) at 1 µg/ml

    Lane 1 : As above
    Lane 2 : Kidney (Human) Tissue Lysate (ab7920) at 20 µg

    Secondary
    Lane 2 : IRDye 680 Conjugated Goat Anti-Mouse IgG (H+L) at 1/10000 dilution


    Predicted band size : 30 kDa
    Observed band size : 30 kDa
  • ICC/IF image of ab28947 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab28947, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • Sandwich ELISA for the detection of NQO1. ab28947 (1/500) was used as the capture antibody. A rabbit polyclonal raised againts the C-terminal end of NQO1 was used for the detection. Please refer to abreview for further experimental details.

    See Abreview

  • Sandwich ELISA for the detection of NQO1, using ab28947 (1/500) as the capture antibody and ab34173 (1/1000) for the detection.
  • Overlay histogram showing HeLa cells stained with ab28947 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab28947, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunohistochemical analysis of dog skin tissue, staining NQO1 with ab28947.

    Tissue was fixed with formaldehyde and antigen retrieval was by heat mediation in a citrate buffer (pH 6). Samples were incubated with primary antibody (1/175 in BSA in TBS) for 45 minutes. ab98784 rabbit polyclonal to anti-mouse HRP (IgG (1/500) was used as the secondary antibody.

    See Abreview

References for Anti-NQO1 antibody [A180] (ab28947)

This product has been referenced in:
  • Kigoshi Y  et al. CACUL1/CAC1 Regulates the Antioxidant Response by Stabilizing Nrf2. Sci Rep 5:12857 (2015). WB ; Human . Read more (PubMed: 26238671) »
  • Schäfer M  et al. Activation of Nrf2 in keratinocytes causes chloracne (MADISH)-like skin disease in mice. EMBO Mol Med 6:442-57 (2014). IHC-P ; Mouse . Read more (PubMed: 24503019) »

See all 25 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (Human colorectal carcinoma)
Permeabilization Yes - Saponin
Gating Strategy Population gathered excluding debris
Specification Human colorectal carcinoma
Preparation Cell harvesting/tissue preparation method: Trypsinisation
Sample buffer: 5% BSA in PBS
Fixation Formaldehyde
Username

Miss. Jasmine Stanley-Ahmed

Verified customer

Submitted Aug 19 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Panc1 tumor)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH=6
Permeabilization No
Specification Panc1 tumor
Blocking step Background Buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 12 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Panc1)
Permeabilization Yes - 0.25% Triton X-100
Specification Panc1
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 12 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rabbit Tissue sections (Kidney)
Specification Kidney
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization No
Username

Abcam user community

Verified customer

Submitted Apr 15 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (Skin)
Specification Skin
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization No
Username

Abcam user community

Verified customer

Submitted Jan 16 2013

Thank you very much for your email.

The antibody should have worked with the described conditions we are unsure about the cause of band at 50 kDa. There could be one or 2 reasons reason

- The stored lysates are used; I would recomm...

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Thank you for contacting us. I am sorry to hear about the wrong band size.

Could you please provide theinformationwhich will help us to understand the cause of wrong band size. The info we need is

- Cell type and name e.g. human he...

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Thank you for taking the time to contact us. I am sorry to hear you have some concerns regarding the results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for the applicati...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Monkey Tissue sections (Liver)
Specification Liver
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: pH 9.0
Permeabilization No
Username

Abcam user community

Verified customer

Submitted Jan 18 2012

I am very pleased to hear you would like to accept our offer and test ab28947 in monkey. This code will give you 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for monkey and include this code in the &...

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