Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) is a rabbit monoclonal antibody detecting Nrf2 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF, Dot Blot. Suitable for Human.
- Biophysical QC for unrivalled batch-batch consistency
- Over 200 publications
- Trusted since 2009
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IHC-P | IP | Dot | WB | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|---|
Human | Tested | Not recommended | Expected | Tested | Tested | Tested |
Synthetic peptide | Not recommended | Not recommended | Tested | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/500 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/5000 - 1/50000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/250 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/80 - 1/100 | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
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The protein expressed by the NFE2L2 gene functions as a transcription factor that plays a key role in responding to oxidative stress by binding to antioxidant response elements (ARE) in the promoter regions of various cytoprotective genes, such as phase 2 detoxifying enzymes, to promote their expression and neutralize reactive electrophiles. Under normal conditions, it is ubiquitinated and degraded in the cytoplasm by the BCR(KEAP1) complex. However, oxidative stress leads to the inhibition of the BCR(KEAP1) complex, allowing NFE2L2/NRF2 to accumulate in the nucleus, form heterodimers with small Maf proteins, and bind to ARE elements. Additionally, the NFE2L2/NRF2 pathway is activated by selective autophagy, where KEAP1 interacts with SQSTM1/p62, inactivating the BCR(KEAP1) complex and leading to nuclear NFE2L2/NRF2 accumulation and cytoprotective gene expression. It may also be involved in activating genes of the beta-globin cluster by facilitating enhancer activity of hypersensitive site 2 of the beta-globin locus control region. This supplementary information is collated from multiple sources and compiled automatically.
NRF2, NFE2L2, Nuclear factor erythroid 2-related factor 2, NF-E2-related factor 2, NFE2-related factor 2, Nrf-2
Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) is a rabbit monoclonal antibody detecting Nrf2 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF, Dot Blot. Suitable for Human.
- Biophysical QC for unrivalled batch-batch consistency
- Over 200 publications
- Trusted since 2009
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Species reactivity
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Blocking buffer: 5% NFDM/TBST, dilution buffer: 5% NFDM /TBST, exposure time: 15 seconds
All lanes: Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) at 1/10000 dilution
Lane 1: Untreated HepG2 (human hepatocellular carcinoma) whole cell lysates 20μg
Lane 2: HepG2 (human hepatocellular carcinoma) treated with Alkaline Phosphatase (AP) whole cell lysates 20μg.
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution
Predicted band size: 68 kDa
Observed band size: 100 kDa
Representative findings of Nrf2 staining in carcinoma (left), in low grade dysplasia (middle), and in non-atypical epithelium (right).
Corresponding PLA signals are displayed in the lower row. Scale bar; 100 μm.
Surgical specimens were transferred to 10% buffered formalin and fixed overnight. The fixed samples were embedded in paraffin, and serially sliced into 5 μm sections. After dewaxing, sections were autoclaved at 120°C for 1 min in 10 mM sodium citrate buffer (pH 6.0), and immersed in 0.3% H2O2. They were then incubated overnight at 4°C with primary antibody to Nrf2 (diluted 1:200). The sections were rinsed with 1×PBS and incubated with the secondary antibody conjugated with horseradish peroxidase at room temperature for 1 hour. The sections were then stained with 3.3′-diaminobenzidinetetrahydrochloride (DAB) and counterstained with hematoxylin.
(A) Human islets were treated with dh404 for 0.5, 1 or 2 hours. The treated and untreated samples were stained with Nrf2 antibody ab76026 (Green) and DAPI (Blue). The con-focal microscope clearly showed that the Nrf2 translocation from cytoplasm to nucleus in the dh404 treated human islet cells
Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified ab76026 at a dilution of 1 in 80 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
B) Healthy skin (top two rows) and KS skin tissue (bottom row) were double-stained for LANA-1 (Alexa-Fluor 594- red) and host phosphorylated pNrf2 (ab76026) (Alexa-Fluor®488 – green). DAPI was used to visualize the nuclei, and the triple merge of LANA-1, pNrf2 and DAPI is shown in the third column.
Yellow square = enlarged area.
Immunofluorescence staining of HepG2 cells with purified ab76026 at a working dilution of 1/100, counter-stained with DAPI. The treated cells were treated with alkaline phosphatase for 1 h at 37°C. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab76026 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) at 1/50000 dilution
Lane 1: untreated HepG2 cell lysate at 10 µg
Lane 2: HepG2 treated with phosphatase lysate at 10 µg
All lanes: HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 68 kDa
Observed band size: 90 kDa
Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab76026 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Dot blot analysis of Nrf2 peptides using unpurified ab76026 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated secondary antibody at 1/1000 dilution. Blocking and diluting buffer was 5% NFDM/TBST.
Lane 1: Nrf2 (pS40) phospho peptide
Lane 2: Nrf2 non-phospho peptide
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using unpurified ab76026 at 1/100 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
All lanes: Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) at 1/20000 dilution
Lane 1: HepG2 cell lysate at 10 µg
Lane 2: HepG2 cell lysate treated with AP at 10 µg
All lanes: HRP labelled goat anti-rabbit at 1/1000 dilution
Predicted band size: 68 kDa
Observed band size: 90 kDa
Unpurified ab76026 staining Nrf2 (phospho S40) in Human normal lung tissue sections by IHC-P (Formaldehyde-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% casein for 30 minutes at 4°C. Antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/50) in 1% casein for 24 hours at 4°C. An undiluted HRP-conjugated Goat polyclonal to rabbit IgG was used as the secondary antibody.
Unpurified ab76026 showing positive staining in Breast carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Unpurified ab76026 showing positive staining in Cervical carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Unpurified ab76026 showing positive staining in Ovarian carcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Unpurified ab76026 showing positive staining in Normal tonsil tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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