Anti-Nrf2 antibody (ab137550)
Key features and details
- Rabbit polyclonal to Nrf2
- Suitable for: IP, ChIP, WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- 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
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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
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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.
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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
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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
Associated products
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Assay kits
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
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.
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| ChIP |
Use at an assay dependent concentration.
Assay dependent |
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| 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.
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| IHC-P | (1) |
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Notes |
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IP
1/100 - 1/500. |
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ChIP
Use at an assay dependent concentration. Assay dependent |
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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. |
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ICC/IF
1/100 - 1/1000. |
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IHC-P
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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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
modificationsPhosphorylation 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
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Database links
- Entrez Gene: 497024 Cow
- Entrez Gene: 4780 Human
- Omim: 600492 Human
- SwissProt: Q5NUA6 Cow
- SwissProt: Q16236 Human
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Alternative names
- erythroid derived 2 antibody
- HEBP1 antibody
- like 2 antibody
see all
Images
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Western blot - Anti-Nrf2 antibody (ab137550)All lanes : Anti-Nrf2 antibody (ab137550) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : NFE2L2 knockout A549 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : Huh7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 85 kDa why is the actual band size different from the predicted?Western blot: Anti-NFE2L2 antibody (ab137550) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab137550 was shown to bind specifically to NFE2L2. A band was observed at 85 kDa in wild-type A549 cell lysates with no signal observed at this size in NFE2L2 knockout cell line ab285359 (knockout cell lysate ab289682). To generate this image, wild-type and NFE2L2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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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.
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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.
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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.
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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.
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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
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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.
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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
Datasheets and documents
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SDS download
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Datasheet download
References (335)
ab137550 has been referenced in 335 publications.
- Tang R et al. Sulforaphane activates anti-inflammatory microglia, modulating stress resilience associated with BDNF transcription. Acta Pharmacol Sin 43:829-839 (2022). PubMed: 34272506
- Chen T et al. Multiple myeloma cells depend on the DDI2/NRF1-mediated proteasome stress response for survival. Blood Adv 6:429-440 (2022). PubMed: 34649278
- Fu R et al. Neuroprotective Effects of Tetrahydroxystilbene Glucoside against Rotenone-Induced Toxicity in PC12 Cells. Biol Pharm Bull 45:143-149 (2022). PubMed: 34707025
- Gwak SY et al. Potential Role of Heme Oxygenase-1 in the Resolution of Experimentally Induced Colitis through Regulation of Macrophage Polarization. Gut Liver 16:246-258 (2022). PubMed: 34737242
- Beà A et al. Cardiac fibroblasts display endurance to ischemia, high ROS control and elevated respiration regulated by the JAK2/STAT pathway. FEBS J 289:2540-2561 (2022). PubMed: 34796659