RabMAb

Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

Overview

  • Product nameAnti-Nrf2 (phospho S40) antibody [EP1809Y]
    See all Nrf2 primary antibodies
  • Description
    Rabbit monoclonal [EP1809Y] to Nrf2 (phospho S40)
  • Tested applicationsICC/IF, IHC-P, WB, IP, ICC, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic phospho-peptide (Human) corresponding to residues near serine 40 of Nrf2.

  • Positive control
    • Human breast carcinoma tissue, HepG2 cell lysate
  • General notes

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

Our Abpromise guarantee covers the use of ab76026 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.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/5000 - 1/50000. Predicted molecular weight: 68 kDa.Can be blocked with Nrf2 (phospho S40) peptide (ab133404).
IP 1/40.
ICC 1/100 - 1/250.
Flow Cyt 1/100.

Target

  • FunctionTranscription 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 specificityWidely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.
  • Sequence similaritiesBelongs to the bZIP family. CNC subfamily.
    Contains 1 bZIP domain.
  • DomainAcidic 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 localizationCytoplasm > 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 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

Anti-Nrf2 (phospho S40) antibody [EP1809Y] images

  • Overlay histogram showing Jurkat cells stained with ab76026 (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 (ab76026, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • All lanes : Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) at 1/20000 dilution

    Lane 1 : HepG2 cell lysate
    Lane 2 : HepG2 cell lysate treated with AP

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP labelled goat anti-rabbit at 1/1000 dilution

    Predicted band size : 68 kDa
    Observed band size : 90 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab76026 at 1/100 dilution.
  • ab76026 staining Nrf2 (phospho S40) in Human normal lung tissue sections by IHC-P (Formaldehyde-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% casein for 30 minutes at 4°C. Antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/50) in 1% casein for 24 hours at 4°C. An undiluted HRP-conjugated Goat polyclonal to rabbit IgG was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab76026 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab76026, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.

  • ab76026 showing positive staining in Breast carcinoma tissue.

  • ab76026 showing positive staining in Cervical carcinoma tissue.

  • ab76026 showing positive staining in Ovarian carcinoma tissue.

  • ab76026 showing positive staining in Normal tonsil tissue.

References for Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

This product has been referenced in:
  • Stachel I  et al. Modulation of nuclear factor E2-related factor-2 (Nrf2) activation by the stress response gene immediate early response-3 (IER3) in colonic epithelial cells: a novel mechanism of cellular adaption to inflammatory stress. J Biol Chem 289:1917-29 (2014). Read more (PubMed: 24311782) »
  • Liu Z  et al. Tanshinone IIA suppresses cholesterol accumulation in human macrophages: role of heme oxygenase-1. J Lipid Res 55:201-13 (2014). Human . Read more (PubMed: 24302760) »

See all 18 Publications for this product

Product Wall

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Tissue lysate - whole (liver)
Specification liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Dec 19 2014



I believe that the Nrf2 that does enter the nucleus is acetylated and phosphorylated, which will make the protein run more slowly through the SDS PAGE gel, making its apparent molecular weight seem larger than based on theoretical amino acid ...

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Thank you for contacting us. The IgG concentration of lot GR37038-5 is 0.114 mg/ml. For a 1 ug/ml working concentration, a dilution of 1/100 will be acceptable, or 1/114 to be more precise. Please let me know if you have any questions or any difficulty...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HepG2 cell line)
Loading amount 30 µg
Specification HepG2 cell line
Gel Running Conditions Reduced Denaturing (10% SDS-PAGE)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 01 2012

Thank you again for contacting us.

I consulted the lab in order to give you the right information relating to your question about the reactivity of our anti phospho Nrf2 antibody (ab76026).
We have inconsistent testing data about the prod...

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Thank you for your reply.

These products are produced in house by our Epitomics group. Our labs use http://www.abcam.com/index.html?pageconfig=resource&rid=14719which areSDS-PAGE Western Blot gels similar to BisTris. Theseare convenient ...

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Thank you for contacting us.

We use SDS-PAGE gels when testing our antibodies in Western blot. I have also seen that the publications referenced also use SDS-PAGE.



According to multiple publications, the expected MW fo...

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Le laboratoire m'a informé que dans certains cas ils observaient également une bande non-spécifique autour des 250 kDa et que cette bande disparaissait en utilisant l'anticorps à des dilutions plus élevées.
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Merci pour ce complément d'information que j'ai transmis à mes collègues du laboratoire.

Je reviendrai vers vous le plus rapidement possible avec, je l'espère, une solution à ce problème.

Merci de nous avoir contactés.

Je vous envoiecomme convenu cet email auquel vous pouvez répondre et qui me sera alors directement attribué.

Merci donc de me faire parvenir l'image du ou des blot(s) obtenus. Pourri...

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