Anti-Nrf2 (phospho S40) antibody [EP1809Y]

• WB

Lab

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/1000 dilution

Predicted band size: 68 kDa

Observed band size: 100 kDa

false

• ICC/IF

PubMed

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

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/

• Flow Cyt (Intra)

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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).

• IHC-P

Unknown

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.

• IHC-P

Unknown

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 (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.

• IHC-P

Unknown

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.

• ICC/IF

Unknown

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 (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with 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 (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

• IHC-P

Unknown

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.

• IHC-P

AbReview16248****

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.

This image is courtesy of an anonymous Abreview

• IHC-P

Unknown

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.

• IHC-P

PubMed

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.

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/

• IHC-P

Unknown

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.

• WB

Lab

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

false

• ICC/IF

PubMed

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.

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/

• WB

Unknown

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

false

• Dot

Unknown

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