Overview

  • Product name
  • Description
    Rabbit polyclonal to Nrf2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, ChIP, IHC-Fr, WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow
  • Immunogen

    Recombinant fragment, corresponding to a region within internal sequence amino acids 108 to 413 of Human Nrf2 (Q16236).

  • Positive control
    • HeLa cells; HeLa whole cell lysate (ab150035); HeLa cytoplasmic lysate; HeLa membrane; HeLa nucleur lysate; HeLa cytoskeleton lysate; NIH-3T3 cell lysate; Human Cal27 xenograft tissue.
  • General notes
    NRF2 is bound to ubiquitin when it is the nucleus so MW would be higher than cytoplasm.

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab137550 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
IP 1/100 - 1/500.
ChIP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. PubMed: 24846271
WB 1/500 - 1/3000.

Detects a band of approximately 110 kDa (predicted molecular weight: 68 kDa).

ICC/IF 1/100 - 1/1000.
IHC-P 1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

  • All lanes : Anti-Nrf2 antibody (ab137550) at 1/1000 dilution

    Lane 1 : Non-transfected (-) 293T whole cell extracts
    Lane 2 : NRF2-transfected (+) 293T whole cell extracts

    Lysates/proteins at 30 µg per lane.
  • ab137550 staining Nrf2 in Rat liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/250 in BSA in TBS) for 30 minutes. A undiluted HRP-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • ab137550 staining Nrf2 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde. Samples were incubated with primary antibody diluted at 1/500 (green). Phalloidin, a cytoskeleton marker, diluted at 1/200 (red) was used.

  • ChIP was performed with HepG2 chromatin extract and 5 μg of either normal rabbit IgG or Anti-Nrf2 antibody (ab137550). The precipitated DNA was detected by PCR with primer set targeting to GCLC gene locus.

  • Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 cells labeling Nrf2 with ab137550 at 1/500 dilution (green). Cells were fixed in 4% paraformaldehyde at RT for 15 min. Hoechst nuclear staining (blue).

  • Anti-Nrf2 antibody (ab137550) at 1/500 dilution + C8D30 whole cell extract at 30 µg
  • ab137550 at 5 μg immunoprecipitating Nrf2 in HepG2 whole cell extracts.

    Lane 1 (-): Ctrl IgG instead of ab137550 in HepG2 whole cell extracts.
    Lane 2 (+): ab137550 + HepG2 whole cell extracts.

    For western blotting an anti-Rabbit IgG was used as a secondary reagent.

  • Immunofluorescence analysis of paraformaldehyde-fixed HeLa, labelling Nrf2 with an137550 at 1/200 dilution.
  • Immunohistochemical analysis of paraffin-embedded Cal27 xenograft, labelling Nrf2 with ab137550 at 1/500 dilution.
  • All lanes : Anti-Nrf2 antibody (ab137550) at 1/1000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HeLa cytoplasmic lysate
    Lane 3 : HeLa nuclear lysate

    Lysates/proteins at 20 µg per lane.

References

This product has been referenced in:
  • Ma YF  et al. Tea polyphenols protect bovine mammary epithelial cells from hydrogen peroxide-induced oxidative damage in vitro by activating NFE2L2/HMOX1 pathways. J Dairy Sci 102:1658-1670 (2019). Read more (PubMed: 30594360) »
  • Zhang L  et al. Sinomenine Attenuates Traumatic Spinal Cord Injury by Suppressing Oxidative Stress and Inflammation via Nrf2 Pathway. Neurochem Res N/A:N/A (2019). Read more (PubMed: 30603983) »
See all 47 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Bos taurus Cell lysate - whole cell (MAC-T cell line)
Gel Running Conditions
Non-reduced Denaturing (10%)
Loading amount
60 µg
Treatment
transfected with shnrf2
Specification
MAC-T cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jun 01 2017

Application
Western blot
Loading amount
25000 cells
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (Prostate)
Specification
Prostate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 29 2014

Answer



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 sequence alone.

As is noted in the Q&A from April 24 2013:
"phosphorylation of Ser-40 by PKC in response to oxidative stress dissociates NFE2L2 from its cytoplasmic inhibitor KEAP1, promoting its translocation into the nucleus by similarity. And acetylation at Lys-596 and Lys-599 increases nuclear localization"

Read More

Answer


The lab got back to me with the following explanations:

The majority of signal in ICC/IF data is on nuclear and less in cytoplasm.
Although NRF2 will translocate into the nucleus by oxidation, this does however not mean that this protein is only expressed in the cytoplasm normally. The cell line tested by ICC/IF is a cancer cell line.

In addition, phosphorylation of Ser-40 by PKC in response to oxidative stress dissociates NFE2L2 from its cytoplasmic inhibitor KEAP1, promoting its translocation into the nucleus by similarity.
And acetylation at Lys-596 and Lys-599 increases nuclear localization whereas deacetylation by SIRT1 enhances cytoplasmic presence.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Liver)
Specification
Liver
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 Citrate
Permeabilization
No

Abcam user community

Verified customer

Submitted Jan 29 2013

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

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