Name: Anti-Nrf2 (phospho S40) antibody [EP1809Y]
SKU: ab76026
Rating: 4 (18 reviews)

Anti-Nrf2 (phospho S40) antibody [EP1809Y] is a rabbit recombinant monoclonal antibody that is used to detect Nrf2 in Dot, Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Human samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity and sensitivity confirmed in immunohistochemistry (IHC) with multi-tissue microarray (TMA) validation
- Cited in over 200 publications
- Trusted since 2009

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Alexa Fluor® 647 Anti-Nrf2 (phospho S40) antibody [EP1809Y]
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Alexa Fluor® 750 Anti-Nrf2 (phospho S40) antibody [EP1809Y]

Images

Western blot - 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-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

Tested
Tested

Species	Dilution info	Notes
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	Dilution info	Notes
Species Synthetic peptide	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Synthetic peptide	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Synthetic peptide	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/5000 - 1/50000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide	Dilution info -	Notes -

Tested
Tested

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.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide	Dilution info -	Notes -

Associated Products

Select an associated product type

8 products for Alternative Product

Target data

See full target information NFE2L2 phospho S40

Function

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.

Alternative names

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

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.
Clone number: EP1809Y
Purification technique: Affinity purification Protein A
Concentration

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

Product Specifications
Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Dot blot, Flow Cyt (Intra), ICC/IF, IHC-P, WB in human samples.
Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) specifically detects Nrf2 Phospho-S40 (UniProt ID: Q16236; Molecular weight: 68kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) has been cited over 208 times in peer reviewed journals and is trusted by the scientific community.
Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) has 16 independent reviews from customers.

Related Products
Antibody clone EP1809Y is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 488, Alexa Fluor® 647, Alexa Fluor® 555, Alexa Fluor® 594, PE, APC, Alexa Fluor® 750 (ab237464, ab237465, ab320134, ab320135, ab320136, ab320137, ab321712).

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:

- 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.

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.

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.

16 product images

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
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.
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)
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).
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)
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)
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)
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)
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)
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)
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
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)
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)
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)
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)
Unpurified ab76026 showing positive staining in Normal tonsil tissue.

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