Overview

  • Product nameAnti-Nrf2 antibody [EP1808Y]
    See all Nrf2 primary antibodies
  • Description
    Rabbit monoclonal [EP1808Y] to Nrf2
  • Tested applicationsSuitable for: ChIP, IHC-Fr, Flow Cyt, ICC/IF, WB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Nrf2 aa 550 to the C-terminus (C terminal). The exact sequence is proprietary.
    (Peptide available as ab167152)

  • Positive control
    • WB: MG-132 treated HeLa whole cell lysate, Saos-2, THP-1 and HepG2 cell lysate. IHC-P: Human pancreatic carcinoma tissue. ICC/IF: HepG2 and HeLa cells. Flow Cyt: HeLa cells. ChIP: Chromatin from HepG2 cells. IP: SAOS-2 whole cell lysate.
  • General notes

    Mouse is not recommended for IHC for weakly staining.

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

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab62352 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
ChIP Use at an assay dependent concentration. PubMed: 22581777
IHC-Fr 1/100. Fix with acetone. Note: antigen retrieval is recommended.
Flow Cyt 1/40.
ICC/IF 1/100 - 1/500.
WB 1/1000 - 1/5000. Predicted molecular weight: 68 kDa.Can be blocked with Nrf2 peptide (ab167152).
IP 1/20.
IHC-P 1/100 - 1/250.

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 (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

Anti-Nrf2 antibody [EP1808Y] images

  • Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Nrf2 with purified ab62352 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. The cells were co-stained with ab195889, an Alexa Fluor® 594-conjugated mouse anti-alpha tubulin antibody (1/200). Nuclei counterstained with DAPI (blue).

    Secondary antibody only control: PBS was used instead of the primary antibody as the negative control.

  • All lanes : Anti-Nrf2 antibody [EP1808Y] (ab62352) at 1/1000 dilution (purified)

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HeLa whole cell lysate treated with MG-132 2 µM for 18 hours
    Lane 3 : Saos-2 whole cell lysate
    Lane 4 : THP-1 whole cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 68 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure:
    Lanes 1 and 2: 15 seconds.
    Lanes 3 and 4: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreatic carcinoma tissue labelling Nrf2 with purified ab62352 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Flow Cytometry analysis of HeLa cells labelling Nrf2 with purified ab62352 at a dilution of 1/60 (red). Cells were fixed with 4% paraformaldehyde. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG (ab172730). Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • ab62352 (purified) at 1/30 immunoprecipitating Nrf2 in SAOS-2 whole cell lysate.

    Lane 1 (input): SAOS-2 whole cell lysate (10µg)
    Lane 2 (+): ab62352 + SAOS-2 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab62352 in SAOS-2 whole cell lysate.

    For western blotting, ab62352 was used at a dilution of 1/500 and ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1000).

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Chromatin was prepared from HepG2 cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab62352 (blue), and 20µl of A/G sepharose bead slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Nrf2 with purified ab62352 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594). DAPI was used to stain the nuclei blue.

    Secondary antibody only control: PBS was used instead of the primary antibody as the negative control.

  • All lanes : Anti-Nrf2 antibody [EP1808Y] (ab62352) at 1/1000 dilution (purified)

    Lane 1 : MEF (Mouse embryonic fibroblast) whole cell lysates
    Lane 2 : MEF (Mouse embryonic fibroblast) treated with MG-132 2uM for 18 hours whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 68 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    Blocking and dilution buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney cancer tissue sections labeling Nrf2 with ab62352 at 1/100 dilution. The tissue was fixed with paraformaldehyde and a heat mediated antigen retrival step was performed with TRIS-EDTA Buffer pH 9.0. Staining with ab62352 at 1/100 was carried out in a dilution buffer with blocking for 30 minutes at 20°C. A undiluted goat anti-rabbit HRP conjugated secondary antibody was used.  

    See Abreview

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Nrf2 with ab62352 at 1/40 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Anti-Nrf2 antibody [EP1808Y] (ab62352) at 1/2000 dilution (unpurified) + HepG2 cell lysate at 10 µg

    Secondary
    goat anti rabbit HRP at 1/2000 dilution

    Predicted band size : 68 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

References for Anti-Nrf2 antibody [EP1808Y] (ab62352)

This product has been referenced in:
  • Latham Birt SH  et al. Endothelial HO-1 induction by model TG-rich lipoproteins is regulated through a NOX4-Nrf2 pathway. J Lipid Res 57:1204-18 (2016). Read more (PubMed: 27185859) »
  • Comerford SA  et al. Hepatoblastoma modeling in mice places Nrf2 within a cancer field established by mutant ß-catenin. JCI Insight 1:e88549 (2016). WB ; Human . Read more (PubMed: 27734029) »

See all 38 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
Sample Human Tissue sections (kidney cancer)
Specification kidney cancer
Permeabilization Yes - Wash Buffer with Tween
Fixative Paraformaldehyde
Username

Mr. Rudolf Jung

Verified customer

Submitted Aug 23 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Bone Marrow derived macrophages (BMDM))
Specification Bone Marrow derived macrophages (BMDM)
Fixative Paraformaldehyde
Permeabilization Yes - 0.2% Triton
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Miss. Sarah Rodrigues Moreira

Verified customer

Submitted Mar 01 2013

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results. We have not had a band reported to use of this size, the predicted MW is around 68 kD but we saw the band a bit higher at 100 kD. We do not have t...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (A549)
Loading amount 15 µg
Specification A549
Treatment 50 uM BSO
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Amelia Romoser

Verified customer

Submitted Jan 28 2011

Application Western blot
Sample Human Cell lysate - whole cell (Human Mammary Epithelial Cells (HMEC))
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 10000 cells
Specification Human Mammary Epithelial Cells (HMEC)
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Aug 22 2016

Application Western blot
Sample Mouse Tissue lysate - whole (Brain tissue)
Gel Running Conditions Reduced Denaturing (10)
Loading amount 50 µg
Specification Brain tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 31 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (brain)
Gel Running Conditions Reduced Denaturing (10)
Loading amount 50 µg
Specification brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 26 2016

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Skin)
Permeabilization No
Specification Skin
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Acetone
Username

Abcam user community

Verified customer

Submitted Nov 03 2015

Application Western blot
Sample Human Cell lysate - whole cell (Placenta)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 30 µg
Specification Placenta
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Vicky

Verified customer

Submitted Sep 30 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Human dermal fibroblasts)
Permeabilization No
Specification Human dermal fibroblasts
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Dr. Kate Hawkins

Verified customer

Submitted Jun 18 2015

1-10 of 30 Abreviews or Q&A

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