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Phospho Nrf2 antibody pS40 [EP1809Y] ab76026 is a rabbit monoclonal antibody that is used in Nrf2 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EP1809Y has been tried and trusted by researchers since 2009 and is cited in >240 publications
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your Nrf2 staining, use in Nrf2 western blotting, IHC, immunofluorescence and flow cytometry

New 20 ul size available


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (AB76026), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (AB76026), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (AB76026), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (AB76026), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (AB76026), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPDotWBICC/IFFlow Cyt (Intra)
Human
Tested
Not recommended
Expected
Tested
Tested
Tested
Synthetic peptide
Not recommended
Not recommended
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1/100 - 1/500

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Synthetic peptide

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Synthetic peptide

Dilution info

-

Notes

-

Tested
Tested

Species

Synthetic peptide

Dilution info

-

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/5000 - 1/50000

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/80 - 1/100

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Not recommended
Not recommended

Species

Synthetic peptide

Dilution info

-

Notes

-

Associated Products

Select an associated product type

8 products for Alternative Product

Target data

Function

Transcription factor that plays a key role in the response to oxidative stress: binds to antioxidant response (ARE) elements present in the promoter region of many cytoprotective genes, such as phase 2 detoxifying enzymes, and promotes their expression, thereby neutralizing reactive electrophiles (PubMed:11035812, PubMed:19489739, PubMed:29018201, PubMed:31398338). In normal conditions, ubiquitinated and degraded in the cytoplasm by the BCR(KEAP1) complex (PubMed:11035812, PubMed:15601839, PubMed:29018201). In response to oxidative stress, electrophile metabolites inhibit activity of the BCR(KEAP1) complex, promoting nuclear accumulation of NFE2L2/NRF2, heterodimerization with one of the small Maf proteins and binding to ARE elements of cytoprotective target genes (PubMed:19489739, PubMed:29590092). The NFE2L2/NRF2 pathway is also activated in response to selective autophagy: autophagy promotes interaction between KEAP1 and SQSTM1/p62 and subsequent inactivation of the BCR(KEAP1) complex, leading to NFE2L2/NRF2 nuclear accumulation and expression of cytoprotective genes (PubMed:20452972). The NFE2L2/NRF2 pathway is also activated during the unfolded protein response (UPR), contributing to redox homeostasis and cell survival following endoplasmic reticulum stress (By similarity). May also 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 (PubMed:7937919). Also plays an important role in the regulation of the innate immune response and antiviral cytosolic DNA sensing. It is a critical regulator of the innate immune response and survival during sepsis by maintaining redox homeostasis and restraint of the dysregulation of pro-inflammatory signaling pathways like MyD88-dependent and -independent and TNF-alpha signaling (By similarity). Suppresses macrophage inflammatory response by blocking pro-inflammatory cytokine transcription and the induction of IL6 (By similarity). Binds to the proximity of pro-inflammatory genes in macrophages and inhibits RNA Pol II recruitment. The inhibition is independent of the NRF2-binding motif and reactive oxygen species level (By similarity). Represses antiviral cytosolic DNA sensing by suppressing the expression of the adapter protein STING1 and decreasing responsiveness to STING1 agonists while increasing susceptibility to infection with DNA viruses (PubMed:30158636). Once activated, limits the release of pro-inflammatory cytokines in response to human coronavirus SARS-CoV-2 infection and to virus-derived ligands through a mechanism that involves inhibition of IRF3 dimerization. Also inhibits both SARS-CoV-2 replication, as well as the replication of several other pathogenic viruses including Herpes Simplex Virus-1 and-2, Vaccinia virus, and Zika virus through a type I interferon (IFN)-independent mechanism (PubMed:33009401).

Alternative names

NRF2, NRF2, NFE2L2, Nuclear factor erythroid 2-related factor 2, NF-E2-related factor 2, NFE2-related factor 2, Nrf-2

Recommended products

Phospho Nrf2 antibody pS40 [EP1809Y] ab76026 is a rabbit monoclonal antibody that is used in Nrf2 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EP1809Y has been tried and trusted by researchers since 2009 and is cited in >240 publications
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your Nrf2 staining, use in Nrf2 western blotting, IHC, immunofluorescence and flow cytometry

New 20 ul size available

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EP1809Y

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Nrf2 also known as nuclear factor erythroid 2-related factor 2 is a transcription factor with a molecular weight of approximately 66 kDa. It plays a mechanical role in regulating the expression of antioxidant proteins that protect against oxidative damage triggered by injury and inflammation. Nrf2 is ubiquitously expressed in various tissues including the liver lungs and skin. Nrf2 activation occurs when it translocates from the cytoplasm to the nucleus to bind the antioxidant response element (ARE) in the DNA starting transcription of target genes.

Biological function summary

Nrf2 acts as an important regulator of the cellular antioxidant response. It works in conjunction with its partner protein Keap1 forming a complex that controls its stability and degradation. Under normal conditions Keap1 keeps Nrf2 in the cytoplasm where it is targeted for ubiquitination and degradation. Once activated by oxidative stress or electrophiles Nrf2 dissociates from Keap1 thereby avoiding degradation and relocates to the nucleus to activate the transcription of ARE-dependent genes. This activity boosts the cellular response to oxidative stress by inducing genes involved in detoxification and cellular defense.

Pathways

Nrf2 plays a significant role in the oxidative stress response and detoxification pathways. Nrf2 activation is linked closely to the PI3K/Akt signaling pathway which influences cell survival growth and metabolism. This pathway also interacts with other important proteins like GSK-3β which can modulate Nrf2 activity and stability. Through these pathways Nrf2 orchestrates a defense mechanism against reactive oxygen species (ROS) by boosting the expression of antioxidant enzymes and detoxifying proteins.

Associated diseases and disorders

Nrf2 has been associated with conditions like cancer and neurodegenerative diseases. In cancer aberrant Nrf2 activation may lead to enhanced tumor survival by increasing expression of cytoprotective genes which makes cancer cells resistant to chemotherapy. Nrf2 also interacts with proteins such as p53 which play roles in tumor suppression and cellular stress responses. In neurodegenerative disorders reduced Nrf2 activity can contribute to oxidative stress leading to neuron damage and disease progression with proteins like amyloid-beta also being linked with oxidative processes affected by Nrf2 functionality.

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:
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16 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail
    Inui et al PLoS One. 2013 Sep 24;8(9):e74398. doi: 10.1371/journal.pone.0074398. eCollection 2013. Fig 2. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    Nrf2 was abundantly expressed in carcinomas, low grade dysplasias, and non-atypical epithelia of oral tissue.

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail
    Masuda et al PLoS One. 2015 Jun 25;10(6):e0131012. doi: 10.1371/journal.pone.0131012. eCollection 2015. Fig 1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    Nrf2 Translocation from cytoplasm to nucleus.

    (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

  • Flow Cytometry (Intracellular) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail
    Giyshi et al PLoS Pathog. 2014 Oct 23;10(10):e1004460. doi: 10.1371/journal.ppat.1004460. eCollection 2014 Oct. Fig 1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    Immunofluorescence analysis of Nrf2 levels in Kaposi's sarcoma skin lesions.

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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.

  • Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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

    Secondary

    All lanes: HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 68 kDa

    Observed band size: 90 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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 - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Dot Blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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.

  • Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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.

    Secondary

    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

  • Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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

    Secondary

    All lanes: HRP labelled goat anti-rabbit at 1/1000 dilution

    Predicted band size: 68 kDa

    Observed band size: 90 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail
    This image is courtesy of an anonymous customer review

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    Unpurified ab76026 showing positive staining in Breast carcinoma tissue.


    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    Unpurified ab76026 showing positive staining in Cervical carcinoma tissue.


    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    Unpurified ab76026 showing positive staining in Ovarian carcinoma tissue.


    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)

    Unpurified ab76026 showing positive staining in Normal tonsil tissue.


    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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