Overview

  • Product name

  • Description

    Mouse monoclonal to Nrf2
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-P, WB, ELISAmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Chimpanzee, Monkey, Gorilla, Orangutan
  • Immunogen

    Recombinant fragment: SPSVASPEHS VESSSYGDTL LGLSDSEVEE LDSAPGSVKQ NGPKTPVHSS GDMVQPLSPS QGQSTHVHDA QCENTPEKEL PVSPGHRKTP FTKDKHSSRL , corresponding to amino acids 351-450 of Human Nrf2 with proprietary tag Q16236

  • Positive control

    • WB: PC12 and Nrf2 transfected 293T cell lysates. ICC/IF: HeLa cells. IHC-P: Human lung tissue.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 15 May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab89443 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 at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Predicted molecular weight: 68 kDa.
ELISA Use at an assay dependent concentration.

Target

  • Function

    Transcription activator that binds to antioxidant response (ARE) elements in the promoter regions of target genes. Important for the coordinated up-regulation of genes in response to oxidative stress. May be involved in the transcriptional activation of genes of the beta-globin cluster by mediating enhancer activity of hypersensitive site 2 of the beta-globin locus control region.
  • Tissue specificity

    Widely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.
  • Sequence similarities

    Belongs to the bZIP family. CNC subfamily.
    Contains 1 bZIP domain.
  • Domain

    Acidic activation domain in the N-terminus, and DNA binding domain in the C-terminus.
  • Post-translational
    modifications

    Phosphorylation of Ser-40 by PKC in response to oxidative stress dissociates NFE2L2 from its cytoplasmic inhibitor KEAP1, promoting its translocation into the nucleus.
  • Cellular localization

    Cytoplasm > cytosol. Nucleus. Cytosolic under unstressed conditions, translocates into the nucleus upon induction by electrophilic agents.
  • Information by UniProt
  • Database links

  • Alternative names

    • erythroid derived 2 antibody
    • HEBP1 antibody
    • like 2 antibody
    • NF E2 related factor 2 antibody
    • NF-E2-related factor 2 antibody
    • NF2L2_HUMAN antibody
    • NFE2 related factor 2 antibody
    • NFE2-related factor 2 antibody
    • Nfe2l2 antibody
    • Nrf 2 antibody
    • NRF2 antibody
    • Nuclear factor (erythroid derived 2) like 2 antibody
    • Nuclear factor antibody
    • nuclear factor erythroid 2 like 2 antibody
    • Nuclear factor erythroid 2 related factor 2 antibody
    • Nuclear factor erythroid 2-related factor 2 antibody
    • Nuclear factor erythroid derived 2 like 2 antibody
    see all

Images

  • Anti-Nrf2 antibody (ab89443) at 1 µg/ml + PC12 cell lysate at 50 µg

    Predicted band size: 68 kDa
    Observed band size: 68 kDa



    This image was generated using the ascites version of the product.

  • ICC/IF image of ab89443 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat 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 (ab89443, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 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.

    This image was generated using the ascites version of the product.

  • IHC image of Nrf2 staining in Human Lung 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 ab89443, 0.5 µg/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.

    This image was generated using the ascites version of the product.

    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.

  • Overlay histogram showing HepG2 cells stained with ab89443 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab89443, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    This image was generated using the ascites version of the product.

  • All lanes : Anti-Nrf2 antibody (ab89443) at 1 µg/ml

    Lane 1 : Nrf2 transfected 293T cell lysate
    Lane 2 : Non-transfected lysate

    Lysates/proteins at 25 µg per lane.

    Predicted band size: 68 kDa
    Observed band size: 68 kDa
    Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.



    This image was generated using the ascites version of the product.

  • Anti-Nrf2 antibody (ab89443) at 1 µg/ml + recombinant immunogen at 0.2 µg

    Predicted band size: 68 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?



    Western blot against tagged recombinant immunogen using ab89443 antibody at 1µg/ml dilution. Predicted band size of immunogen is 11kDa.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:

  • Li Z  et al. Protective effects of tetramethylpyrazine analogue Z-11 on cerebral ischemia reperfusion injury. Eur J Pharmacol 844:156-164 (2019). Read more (PubMed: 30502344) »
  • Zhou R  et al. Salvianolic acid B, an antioxidant derived from Salvia militarize, protects mice against ?-radiation-induced damage through Nrf2/Bach1. Mol Med Rep 19:1309-1317 (2019). Read more (PubMed: 30535483) »
See all 22 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (BV2 microglia)
Gel Running Conditions
Reduced Denaturing
Loading amount
100 µg
Treatment
5µM LPA for different time points
Specification
BV2 microglia
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Lisha Joshi

Verified customer

Submitted Jun 12 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric buffer PH=6
Specification
brain
Blocking step
BSA 5%, NGS 3%, TRITON 0.2% as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
with no fixation

Mr. guy keller

Verified customer

Submitted Sep 06 2018

Answer

Thank you for contacting us. I am sorry to hear that you have been experiencing problems with antibodies.

The antibody ab89443 was characterized in WB using PC12 cell line lysates. Other possible cell line lysates are HepG2, A-431, HEK293, U-2OS or U251-MG cell line lysates.

ab91421, Hela cell or NIH-3T3 cell line lysates could be used in WB.

For IHC please click the link; http://www.proteinatlas.org/ENSG00000114026

Once you have selected the cell line lysates then please check the product on our website or send me an email, I will then suggest you catalogue numbers for buying.

If however you are performing IHC then please click the link below to see which tissue has high expression and can be used as positive control.

http://www.proteinatlas.org/ENSG00000116044
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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DISCOUNT CODE: ***
Expiration date: September 15th, 2012

I am very pleased to hear you would like to accept our offer and test ab53019 inIP. This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for IP and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

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Es freut mich, dass es geklappt hat!

Wir freuen uns auf eine weitere Zusammenarbeit mit Ihnen!

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Vielen Dank für Ihre Anfrage. Ihre Gutschriftnummer lautet CGBxxx und die zugehörige interne Referenznummer ist y. Diese Daten sollten Ihnen per Post zugegangen sein. Falls dies nicht geschehen ist, tut es uns sehr leid und wir möchten uns dafür entschuldigen. Ich habe Ihnen die Gutschrift auch noch einmal als pdf-Datei angehangen. Falls Sie diese Gutschrift mit einer neuen Bestellung einlösen möchten, geben Sie die Gutschriftnummer bitte bei der Bestellung mit an. Sie wird dann von unserer Finanzabteilung abgeglichen und gegengerechnet. Für weitere Fragen steht Ihnen auch gerne unser Kundendienst (030 896 779 154) beziehungsweise unsere Finanzabteilung (creditcontrol@abcam.com) zur Verfügung. Ich möchte mich in diesem Zusammenhang auch dafür bedanken, dass Sie an unserer Umfrage teilgenommen haben. Ihre konstruktive Kritik, auf dem Datenblatt nicht Westernblots mit unterschiedlichen Molekulargewichten zu veröffentlichen und gegebenenfalls genauere Informationen zu geben, war für uns sehr wichtig. Wir versuchen immer, alle Informationen, die wir über ein Produkt haben, auf unseren Datenblättern anzugeben und zu aktualisieren, sobald wir neue erhalten. Es tut uns leid, dass dies im Fall des Nrf2-Antikörpers ab89443 zu wünschen übrig ließ. Die für ab89443 gezeigten Blots stammen aus dem Labor, das den Antikörper herstellt, und wir würden lieber Daten "aus dem Alltag" zeigen. Aus diesem Grund haben wir das Abreviewprogramm ins Leben gerufen: Wir möchten unsere Kunden immer ermutigen, uns Ihre positiven (und auch negativen!) Ergebnisse mitzuteilen. Wir machen diese Informationen für alle Kunden zugänglich und belohnen diese „Abreviews“ mit Punkten, die gegen Rabatte beim Kauf weiterer Produkte oder Geschenkgutscheine wie z.B. Amazon.com-Gutscheine eingetauscht werden können. Unter der folgenden URL können Sie mehr über unser Abreview-System zu erfahren: https://www.abcam.com/abreviews Falls Sie also irgendwann einmal zu einem Antikörper einen (Bild-)Beitrag liefern möchten, möchten wir Sie einladen unser Abreview-System zu verwenden. Sie können die Rezension durch einen Link direkt auf unserem Produktdatenblatt abschicken. Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Vielen Dank für Ihre Antwort. Es tut mir leid zu hören, dass die Vorschläge in diesem Fall nicht zu einer Verbesserung Ihrer Ergebnisse geführt haben.  Die Referenznummer Ihrer Gutschrift ist CN xxxxx. Es tut mir leid, dass der Antikörper nicht so funktioniert hat, wie auf dem Datenblatt angegeben. Ich habe unsere Finanzabteilung beauftragt, eine Gutschrift für Sie auszustellen. Sie können diese Gutschrift auf folgende Weise verwenden: (1) Gegen die Rechnung dieses Antikörpers verwenden falls diese noch nicht bezahlt wurde (2) Aufbewahren und gegen eine zukünftige Rechnung verwenden (3) Wir stellen Ihne gerne eine komplette Rückerstattung aus, wenn keine offenen Rechnungen mehr ausstehen Um eine Rückerstattung des Kaufpreises zu erhalten oder um zu bestätigen wie Sie diese Gutschrift benutzen möchten, möchte ich Sie bitten, sich mit unserer Finanzabteilung in Verbindung zu setzen. Sie können unsere Finanzabteilung per Email unter der Addresse creditcontrol@abcam.com erreichen. Bitte geben Sie die Referenznummer Ihrer Gutschrift immer mit an. Die Referenznummer ist für Ihre Unterlagen bestimmt. Wir schicken Ihnen die eigentliche Gutschrift per Post oder eMail zu. Sie enthält auch die Gutschriftnummer, die mit dem Buchstaben CGB beginnt. Ich hoffe, dass Sie trotz dieser schlechten Erfahrung weiterhin Kunde bei uns bleiben. Bitte zögern Sie nicht, sich wieder an uns zu wenden, falls Sie weitere Fragen haben.  

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