Recombinant Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1808Y] to Nrf2 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB, ChIP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free
See all Nrf2 primary antibodies -
Description
Rabbit monoclonal [EP1808Y] to Nrf2 - BSA and Azide free -
Host species
Rabbit -
Specificity
The expression of Nrf2 is stimulated by oxidative stress, electrophiles and chemical activators (PMID: 25761198, PMID: 27638861 and PMID: 28587109). Nrf2 antibody (ab62352) detects no signal in most untreated samples in WB. Stimuli treated samples are recommended. We do not recommend using this product in western blot with tissue lysates, however some customers have used this antibody successfully using concentrated samples (see submitted abreviews).
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Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB, ChIPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab167152) -
Positive control
- WB: MG-132 treated HeLa whole cell lysate, Saos-2, THP-1, MG-132 treated HepG2 whole cell lysate, MG-132 treated HCT-116 whole cell lysate. IHC-P: Human pancreatic carcinoma and kidney cancer tissues. ICC/IF: HepG2 and HeLa cells. Flow Cyt (intra): HeLa cells. IP: SAOS-2 whole cell lysate. ChIP: Chromatin prepared from HepG2 cells.
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General notes
ab180845 is the carrier-free version of ab62352.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1808Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-Nrf2 antibody [EP1808Y] (ab194984)
- Alexa Fluor® 647 Anti-Nrf2 antibody [EP1808Y] (ab194985)
- HRP Anti-Nrf2 antibody [EP1808Y] (ab194986)
- Alexa Fluor® 594 Anti-Nrf2 antibody [EP1808Y] (ab206890)
- PE Anti-Nrf2 antibody [EP1808Y] (ab223926)
- APC Anti-Nrf2 antibody [EP1808Y] (ab223927)
- Anti-Nrf2 antibody [EP1808Y] - ChIP Grade (ab62352)
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab180845 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 68 kDa.
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ChIP |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 68 kDa. |
ChIP
Use at an assay dependent concentration. |
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: 4780 Human
- Omim: 600492 Human
- 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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All lanes : Anti-Nrf2 antibody [EP1808Y] - ChIP Grade (ab62352) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate treated with MG-132 2uM for 18h
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 68 kDa
Observed band size: 955-110 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsThis data was developed using ab62352, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concnetration: 5% NFDM/TBST.
ab181602 was used as GAPDH loading control.
The two bands are different isoforms of Nrf2, the molecular weight observed is consistent with what has been described in the literature: PMID: 17512459, PMID: 22703241.
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All lanes : Anti-Nrf2 antibody [EP1808Y] - ChIP Grade (ab62352) at 1/500 dilution
Lane 1 : Wild-type HeLa control MG132 (0 uM, 18 h) cell lysate
Lane 2 : Wild-type HeLa treated MG132 (2 uM, 18 h) cell lysate
Lane 3 : NFE2L2 knockout HeLa control MG132 (0 uM, 18 h) cell lysate
Lane 4 : NFE2L2 knockout HeLa treated MG132 (2 uM, 18 h) cell lysate
Lane 5 : HCT 116 cell lysate
Lane 6 : Saos-2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Nrf2 antibody [EP1808Y] - ChIP Grade staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab62352 was shown to bind specifically to Nrf2. A band was observed at 85 kDa in wild-type HeLa cell lysates with no signal observed at this size in NFE2L2 CRISPR-Cas9 edited cell line ab262507 (CRISPR-Cas9 edited cell lysate ab263934). The band observed in the CRISPR-Cas9 edited lysate lane below 85 kDa is likely to represent a truncated form of Nrf2. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and NFE2L2 CRISPR-Cas9 edited HeLa 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 647). Please refer to ab194985 for protocol details.
ab194985 staining Nrf2 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab194985 at a working dilution 1/100 (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
This product gave a positive signal in 4% formaldehyde (10 min) fixed HeLa cells under the same testing conditions.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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All lanes : Anti-Nrf2 antibody [EP1808Y] - ChIP Grade (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 kDaThis data was developed using ab62352, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure:
Lanes 1 and 2: 15 seconds.
Lanes 3 and 4: 3 minutes. -
Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (PE). Please refer to ab223926 for protocol details.
Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab223926 (red line). The cells were fixed with 4% formaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Triton X-100 for 15 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab223926, 1/5000 dilution) for 30 minutes at 22°C.
Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.
This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Triton X-100 for 15 minutes used under the same conditions.
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 488). Please refer to ab194984 for protocol details.
ab194984 staining Nrf2 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab194984 at a working dilution of 1/100 (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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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 VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Blocking and dilution buffer: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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Intracellular 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
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Intracellular 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (13)
ab180845 has been referenced in 13 publications.
- Chhunchha B et al. Switching of Redox Signaling by Prdx6 Expression Decides Cellular Fate by Hormetic Phenomena Involving Nrf2 and Reactive Oxygen Species. Cells 11:N/A (2022). PubMed: 35455944
- Bu X et al. CD147 confers temozolomide resistance of glioma cells via the regulation of β-TrCP/Nrf2 pathway. Int J Biol Sci 17:3013-3023 (2021). PubMed: 34421346
- Chhunchha B et al. Sulforaphane-Induced Klf9/Prdx6 Axis Acts as a Molecular Switch to Control Redox Signaling and Determines Fate of Cells. Cells 8:N/A (2019). PubMed: 31569690
- Hu T et al. Clinicopathologic significance of CXCR4 and Nrf2 in colorectal cancer. J Biomed Res 27:283-90 (2013). IHC ; Human . PubMed: 23885267
- Ma J et al. PALB2 interacts with KEAP1 to promote NRF2 nuclear accumulation and function. Mol Cell Biol 32:1506-17 (2012). WB . PubMed: 22331464