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    products/primary-antibodies/nrf2-antibody-ab137550.pdf

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Epigenetics and Nuclear Signaling Transcription Other factors
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Anti-Nrf2 antibody (ab137550)

  • Datasheet
  • SDS
Reviews (7)Q&A (2)References (249)

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Western blot - Anti-Nrf2 antibody (ab137550)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody (ab137550)
  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody (ab137550)
  • ChIP - Anti-Nrf2 antibody (ab137550)
  • Western blot - Anti-Nrf2 antibody (ab137550)
  • Immunoprecipitation - Anti-Nrf2 antibody (ab137550)
  • Western blot - Anti-Nrf2 antibody (ab137550)

Key features and details

  • Rabbit polyclonal to Nrf2
  • Suitable for: IP, ChIP, WB, ICC/IF, IHC-P
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Nrf2 antibody [EP1808Y] - ChIP Grade (ab62352)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Nrf2 antibody
    See all Nrf2 primary antibodies
  • Description

    Rabbit polyclonal to Nrf2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, ChIP, WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Cow
  • Immunogen

    Recombinant fragment corresponding to Human Nrf2 aa 108-413 (internal sequence).
    Database link: Q16236

  • Positive control

    • WB: MDA-MB-231 nuclear extract treated with 30 µM tBHQ for 4 hours (+). NRF2-transfected (+, including 3xFlag-tag) and HEK-293T whole cell extracts. ICC/IF: HeLa cells. IHC: Human breast carcinoma tissue. ChIP: HepG2 chromatin extract. IP: HepG2 whole cell extracts.
  • General notes

    NRF2 is bound to ubiquitin when it is the nucleus so MW would be higher than cytoplasm.

    Please note: 

    Nrf2 antibody (ab137550) detects no signal in most untreated samples for WB. Stimuli treated samples are recommended. Nrf2 expression is stimulated by oxidative stress, electrophiles and chemical activators (PMID: 25761198, PMID: 27638861 and PMID: 28587109). 

    WB protocol info included in legend for MDA-MB-231 nuclear extract treated with 30 µM tBHQ for 4 hours (+).

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.00
    Preservative: 0.025% Proclin 300
    Constituents: 78% PBS, 1% BSA, 20% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cardiovascular
    • Heart
    • Cardiac metabolism
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial Biogenesis
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Nucleotide metabolism
    • Molecular processes
    • Mitochondrial transcription
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Signal Transduction
    • Antibodies
    • nrf2
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Assay kits

    • Nrf2 Transcription Factor Assay Kit (Colorimetric) (ab207223)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • NIH/3T3 whole cell lysate (ab7179)
  • Recombinant Protein

    • Recombinant Human Nrf2 protein (ab132356)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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.

Assay dependent

WB (6)
1/500 - 1/3000.

Detects a band of approximately 110 kDa (predicted molecular weight: 68 kDa). NRF2 is bound to ubiquitin when it is the nucleus so MW would be higher than cytoplasm.

Stimuli treated samples are recommended.

Expression levels may be better for nuclear fractions or for cell lines treated with MG132.

ICC/IF
1/100 - 1/1000.
IHC-P (1)
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
IP
1/100 - 1/500.
ChIP
Use at an assay dependent concentration.

Assay dependent

WB
1/500 - 1/3000.

Detects a band of approximately 110 kDa (predicted molecular weight: 68 kDa). NRF2 is bound to ubiquitin when it is the nucleus so MW would be higher than cytoplasm.

Stimuli treated samples are recommended.

Expression levels may be better for nuclear fractions or for cell lines treated with MG132.

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.
  • Target information above from: UniProt accession Q16236 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 497024 Cow
    • Entrez Gene: 4780 Human
    • Omim: 600492 Human
    • SwissProt: Q5NUA6 Cow
    • SwissProt: Q16236 Human
    • 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

    • Western blot - Anti-Nrf2 antibody (ab137550)
      Western blot - Anti-Nrf2 antibody (ab137550)
      All lanes : Anti-Nrf2 antibody (ab137550) at 1/500 dilution

      Lane 1 : Untreated HepG2 whole cell extracts
      Lane 2 : Treated HepG2 whole cell extracts

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : A HRP-conjugated anti-rabbit IgG antibody

      Observed band size: 110 kDa why is the actual band size different from the predicted?



      The observed MW (110kDa) is based on PMID: 22703241

      Various whole cell extracts (30 μg) were separated by 7.5% SDS-PAGE

      Detection: Trident ECL plus-Enhanced.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody (ab137550)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody (ab137550)

      Paraffin embedded human breast carcinoma tissue stained for NRF2 using ab137550 at 1/500 dilution in immunohistochemical analysis.

    • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody (ab137550)
      Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody (ab137550)

      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 1/100 (red) was used.

    • ChIP - Anti-Nrf2 antibody (ab137550)
      ChIP - Anti-Nrf2 antibody (ab137550)

      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.

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

      Lane 1 : Untreated (-) MDA-MB-231 nuclear extract
      Lane 2 : MDA-MB-231 nuclear extract treated with 30 µM tBHQ for 4 hours (+)

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution

      Developed using the ECL technique.

      Observed band size: 110 kDa why is the actual band size different from the predicted?



      7.5% gel.

      Running condition: 80V, 15min; 140V, 40min.

      Transfer condition:Semi-dry, 18 V, 60min (Nitrocellulose membrane)

      Blocking condition: 5% non-fat milk in TBST, RT, 60min.

      Primary antibody incubation: 4°C overnight.

      Secondary antibody incubation: Room temperature for 1 hour.

      Washing condition: 5 ml TBST, 4 x 5min.

      ECL detection. 

      The observed MW (110kDa) is based on PMID: 22703241

    • Immunoprecipitation - Anti-Nrf2 antibody (ab137550)
      Immunoprecipitation - Anti-Nrf2 antibody (ab137550)

      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.

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

      Lane 1 : Non-transfected (-) HEK-293T whole cell extracts
      Lane 2 : NRF2-transfected (+, including 3xFlag-tag) HEK-293T whole cell extracts

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : HRP-conjugated anti-rabbit IgG antibody

      Observed band size: 110 kDa why is the actual band size different from the predicted?



      5% SDS-PAGE

      The observed MW (110kDa) is based on PMID: 22703241

    Protocols

    • ChIP protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (249)

    Publishing research using ab137550? Please let us know so that we can cite the reference in this datasheet.

    ab137550 has been referenced in 249 publications.

    • Chen M  et al. Upregulation Sestrin2 protects against hydrogen peroxide-induced oxidative damage bovine mammary epithelial cells via a Keap1-Nrf2/ARE pathway. J Cell Physiol 236:392-404 (2021). PubMed: 32519422
    • Chen Q  et al. Shenyan Kangfu tablet alleviates diabetic kidney disease through attenuating inflammation and modulating the gut microbiota. J Nat Med 75:84-98 (2021). PubMed: 32997272
    • Martinez R  et al. Mechanistic Analysis of an Extracellular Signal-Regulated Kinase 2-Interacting Compound that Inhibits Mutant BRAF-Expressing Melanoma Cells by Inducing Oxidative Stress. J Pharmacol Exp Ther 376:84-97 (2021). PubMed: 33109619
    • Luo M & Luo Y Imperatorin Relieved Ulcerative Colitis by Regulating the Nrf-2/ARE/HO-1 Pathway in Rats. Inflammation 44:558-569 (2021). PubMed: 33098052
    • Cheng Y  et al. Covalent modification of Keap1 at Cys77 and Cys434 by pubescenoside a suppresses oxidative stress-induced NLRP3 inflammasome activation in myocardial ischemia-reperfusion injury. Theranostics 11:861-877 (2021). PubMed: 33391509
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-9 of 9 Abreviews or Q&A

    Western blot abreview for Anti-Nrf2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - nuclear (Cardiomyocytes)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    5 µg
    Specification
    Cardiomyocytes
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jul 08 2022

    Western blot abreview for Anti-Nrf2 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Dopaminergic Neurons)
    Gel Running Conditions
    Reduced Denaturing (NuPage 4-12% MES buffer)
    Loading amount
    30 µg
    Specification
    Dopaminergic Neurons
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 20 2020

    Western blot abreview for Anti-Nrf2 antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (colon cancer)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    colon cancer
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 13 2020

    Western blot abreview for Anti-Nrf2 antibody

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (rat alveolar epithelial cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Treatment
    5% cigarette smoke extract
    Specification
    rat alveolar epithelial cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 20 2020

    Western blot abreview for Anti-Nrf2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 01 2017

    Western blot abreview for Anti-Nrf2 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jul 29 2014

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nrf2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 29 2013

    Question

    You said "a lack of ubiquitination leads to accumulation of Nrf2 in the nucleus, leading to activation of Nrf2 dependent genes". This makes sense to me, but it still does not explain why, in the nuclear compartment, only the 100 kDa bands shows up. Is there an explanation for this, or is this still not clearly known?

    Read More

    Abcam community

    Verified customer

    Asked on Jul 09 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

    Jeremy Kasanov

    Abcam Scientific Support

    Answered on Jul 09 2014

    Question

    in ICC/IF image: were Hela cells treated as to see nuclear staining?
    can cytoplasmic staining be seen when untreated?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 24 2013

    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 Scientific Support

    Answered on Apr 24 2013

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