
Anti-Nrf2 antibody [EP1808Y] - C-terminal (ab62352)
- Datasheet
- References (130)
- Protocols
Overview
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Product nameAnti-Nrf2 antibody [EP1808Y] - C-terminal
See all Nrf2 primary antibodies -
DescriptionRabbit monoclonal [EP1808Y] to Nrf2 - C-terminal
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Host speciesRabbit
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Tested applicationsSuitable for: ChIP, IHC-Fr, Flow Cyt, ICC/IF, WB, IP, IHC-Pmore details
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Species reactivityReacts with: Mouse, Human
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Immunogen
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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.
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General notes
Mouse is not recommended for IHC (weak staining).
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
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
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEP1808Y
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IsotypeIgG
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Research areas
Associated products
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Alternative Products
- Anti-Nrf2 antibody (ab137550)
- Anti-Nrf2 (phospho S40) antibody [EP1809Y] - BSA and Azide free (ab180844)
- Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
- Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 488) (ab194984)
- Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 647) (ab194985)
- Anti-Nrf2 antibody [EP1808Y] (Phycoerythrin) (ab223926)
- Anti-Nrf2 antibody (ab31163)
- Anti-Nrf2 antibody (ab71890)
- Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)
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Alternative Versions
- Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
- Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 488) (ab194984)
- Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 647) (ab194985)
- Anti-Nrf2 antibody [EP1808Y] (HRP) (ab194986)
- Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 594) (ab206890)
- Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 405) (ab206891)
- Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 568) (ab206893)
- Anti-Nrf2 antibody [EP1808Y] (Phycoerythrin) (ab223926)
- Anti-Nrf2 antibody [EP1808Y] (Allophycocyanin) (ab223927)
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Assay kits
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ChIP Related Products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Related Products
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 |
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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
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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.
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Tissue specificityWidely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.
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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.
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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 localizationCytoplasm > 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: 4780 Human
- Entrez Gene: 18024 Mouse
- Omim: 600492 Human
- SwissProt: Q16236 Human
- SwissProt: Q60795 Mouse
- Unigene: 744006 Human
- Unigene: 1025 Mouse
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Alternative names
- erythroid derived 2 antibody
- HEBP1 antibody
- like 2 antibody
see all
Images
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All lanes : Anti-Nrf2 antibody [EP1808Y] - C-terminal (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
All lanes : 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. -
All lanes : Anti-Nrf2 antibody [EP1808Y] - C-terminal (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
All lanes : 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 minutesBlocking and dilution buffer: 5% NFDM/TBST
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Anti-Nrf2 antibody [EP1808Y] - C-terminal (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? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody [EP1808Y] - C-terminal (ab62352)
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). Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody [EP1808Y] - C-terminal (ab62352)This image is courtesy of an abreview submitted by Rudolf Jung.
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.
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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.
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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. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody (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.
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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).
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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.
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Flow Cytometry analysis of HeLa cells labelling Nrf2 with purified ab62352 at a dilution of 1/60 (red). Cells were fixed with 4% paraformaldehyde. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody (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.
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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.
Protocols
References
This product has been referenced in:
- Li B et al. MIR-144-mediated NRF2 gene silencing inhibits fetal hemoglobin expression in sickle cell disease. Exp Hematol 70:85-96.e5 (2019). Read more (PubMed: 30412705) »
- Narzt MS et al. A novel role for NUPR1 in the keratinocyte stress response to UV oxidized phospholipids. Redox Biol 20:467-482 (2019). Read more (PubMed: 30466060) »