Overview

  • Product name
  • Description
    Goat polyclonal to NQO1
  • Host species
    Goat
  • Specificity
    This products is expected to recognize all three reported isoforms (NP_000894.1; NP_001020604.1; NP_001020605.1)
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-FoFr, ELISA, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
    Predicted to work with: Pig
  • Immunogen

    Synthetic peptide: SIPTDNQIKARK, corresponding to C terminal amino acids 267-274 of Human NQO1.

  • Positive control
    • Recombinant Human NQO1 protein (ab87692) can be used as a positive control in WB. Human kidney, lung and liver lysate
  • General notes
    Principal Names – NQO1; NAD(P)H dehydrogenase, quinone 1; DTD; QR1; DHQU; DIA4; NMOR1; NMORI; diaphorase-4; diaphorase (NADH/NADPH); diaphorase (NADH/NADPH) (cytochrome b-5 reductase); NAD(P)H:menadione oxidoreductase 1, dioxin-inducible 1. Official Gene Symbol - NQO1. GenBank Accession Number – NP_000894. LocusLink ID - 1728. Gene Ontology terms - cytoplasm; xenobiotic metabolism; detoxification response; cytochrome b5 reductase; nitric oxide biosynthesis; NAD(P)H dehydrogenase (quinone); synaptic transmission, cholinergic.

Properties

Applications

Our Abpromise guarantee covers the use of ab2346 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 0.03 - 0.1 µg/ml. Detects a band of approximately 26-30 kDa.Can be blocked with Human NQO1 peptide (ab22904).
IHC-FoFr 1/250. PubMed: 19186165
ELISA 1/64000.
IHC-Fr Use at an assay dependent dilution. PubMed: 7715346

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

  • This image was kindly supplied as part of the review submitted by Lauren Aleksunes. Various mouse liver cytosol samples and human recombinant protein were run on the gel and then incubated with ab2346. This image was kindly supplied as part of the review submitted by Lauren Aleksunes. Various mouse liver cytosol samples and human recombinant protein were run on the gel and then incubated with ab2346.
  • Anti-NQO1 antibody (ab2346) at 0.03 µg/ml + Human Kidney lysate at 35 µg

    Observed band size: 30 kDa
    why is the actual band size different from the predicted?



    Primary incubation was 1 hour. Detected by chemiluminescence.
  • ICC/IF image of ab2346 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2346, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-goat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Lv W  et al. Schwann Cell Plasticity is Regulated by a Weakened Intrinsic Antioxidant Defense System in Acute Peripheral Nerve Injury. Neuroscience 382:1-13 (2018). Read more (PubMed: 29684504) »
  • Prestigiacomo V & Suter-Dick L Nrf2 protects stellate cells from Smad-dependent cell activation. PLoS One 13:e0201044 (2018). Read more (PubMed: 30028880) »
See all 56 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Question
Answer

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Unseres Wissens haben wir noch keine Abbildung von ab2346 Anti-NQO1 antibody in IHC-Fr. Falls Sie uns eine Abbildung Ihrer Ergebnisse mit ab2346 zukommen lassen, kann ich Ihnen zurzeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat und einer Abbildung zusenden. Der Rabatt würde den ganzen Wert von ab2346 abdecken, und gegen eine erneute Bestellung von uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte, dass Sie ab2346 kaufen möchten und bereit sind uns eine Abbildung Ihrer Ergebnisse zukommen zu lassen.

2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper in IHC-Fr.

5.) Senden Sie uns die Abbildung Ihres Ergebnisses mittels eines Abreviews zu und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv. Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

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Question
Answer

Thank you for contacting us. I am sorry that ab2346 is giving no signal in WB on rat samples. Based on the protocol you provided, I would recommend trying a primary antibody dilution of 1:1000. Depending on the expression of NQO1 in your samples, you may need to use as much as a 1:500 dilution.

Was the WB run under denaturing conditions? This antibody has been optimized under reduced / denaturing conditions and I would recommend using a lysis buffer to prepare your samples, such as:

Tris-Triton buffer: (Cytoskeletal proteins)

10 mM Tris, pH 7.4
100 mM NaCl
1 mM EDTA
1 mM EGTA
1% Triton X-100
10% glycerol
0.1% SDS
0.5% deoxycholate

You also seem to be loading a very large amount of protein. For most western blots, I would recommend loading between 20-40ug per lane. At higher concentrations, you can often observe inefficient running and transferring. The high protein concentration can also sometimes lead to an inverse banding effect where you see lighter bands on a darker background.

I hope this helps, if not, please let me know and I will be happy to help you further.

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Answer

I'm very sorry to hear you are experiencing problems with lot 105593, we do not have any more vials of lot 44897 or 105593, and if you could provide me with the order details of the vials of lot 105593 I can certainly arrange replacement vials if you have purchased those in the last 90 days, I look forward to hearing from you to arrange this,

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Question

DESCRIPTION OF THE PROBLEM I ran liver cytosol from NQO1 knockout mice (obtained from the NIH) and had the same bands in the wild-type and knockout mice. This leads us to believe that we are not detecting mouse NQO1 using ab2346. Additionally, a prototypical inducer of NQO1 (BHA) was given to mice. Similar to the knockout mice, there was no difference in bands detected. While we are primarily interested in mouse NQO1, the antibody did recognize human recombinant NQO1. SAMPLE CD-1 mouse liver cytosol, NQO1 knockout mouse liver cytosol, liver cytosol from BHA-treated mice PRIMARY ANTIBODY ab2346 (anti-mouse NQO1) 2 ug/ml in TBS-T for 2 hours SECONDARY ANTIBODY 1:30,000 anti-goat IgG (NB 710-H) for 20 minutes DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Positive control - human recombinant NQO1 Sigma Chemicals, D-1315 - WORKED Positive control - butylated hydroxyanisole (BHA) treated mice - DID NOT WORK Negative control - NQO1 knockout mouse liver cytosol from NIH - DID NOT WORK ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION Samples prepared in sucrose-Tris buffer containing aprotonin and PMSF. Samples were boiled 3 minutes prior to electrophoresis AMOUNT OF PROTEIN LOADED 50 ug protein/well for cytosol samples 4 ug protein/well for recombinant human NQO1 (Sigma Chemicals, D-1315) ELECTROPHORESIS/GEL CONDITIONS SDS-PAGE 10% running/4% stacking gel TRANSFER AND BLOCKING CONDITIONS Transfer in NOVEX system, overnight at 4 C onto PVDF membrane, transfer assessed by Coomassie staining at end of immunostaining, blocked in 1.5% fish gelatin as per Novus Biologicals HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? We have been using knockout liver and are NOT seeing any difference in bands compared to wildtype tissue suggesting that ab2346 is not detecting mouse NQO1

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Answer

I'm sorry to hear that you are having problems with this antibody. I have contacted the originator of ab2346 and there is a new batch of this antibody with better ELISA results than the previous batch which you received. The new batch has only been tested so far on human samples (human kidney and liver lysate) but is expected to cross-react with mouse due to sequence homology. Would you like a replacement vial of the new batch free of charge? Did you order directly from Abcam? Please send me your Abcam order# or PO# and I can send you the replacement vial once it comes into stock.

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Question
Answer

This antibody has only been tested on mouse liver lysates in Western blot.

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