Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)
- Product nameAnti-Nrf2 (phospho S40) antibody [EP1809Y]See all Nrf2 primary antibodies ...
- DescriptionRabbit monoclonal [EP1809Y] to Nrf2 (phospho S40)
- Tested applicationsICC/IF, IHC-P, WB, IP, ICC, Flow Cyt more details
- Species reactivityReacts with: Human
Does not react withMouse, Rat
Synthetic phospho-peptide (Human) corresponding to residues near serine 40 of Nrf2.
- Positive control
- Human breast carcinoma tissue, HepG2 cell lysate
- General notesProduced under U.S. Patent No. 5,675,063.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
- PurityTissue culture supernatant
- Clonality Monoclonal
- Clone numberEP1809Y
- Pathways and Processes
- Metabolic signaling pathways
- Nucleotide metabolism
- Molecular processes
- Mitochondrial transcription
Our Abpromise guarantee covers the use of ab76026 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||ICC/IF: Use a concentration of 1 µg/ml.|
|IHC-P||IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||WB: 1/5000 - 1/50000. Predicted molecular weight: 68 kDa.Can be blocked with Nrf2 (phospho S40) peptide (ab133404).|
|ICC||ICC: 1/100 - 1/250.|
|Flow Cyt||Flow Cyt: 1/100.|
- FunctionTranscription 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 specificityWidely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.
- Sequence similaritiesBelongs to the bZIP family. CNC subfamily.
Contains 1 bZIP domain.
- DomainAcidic activation domain in the N-terminus, and DNA binding domain in the C-terminus.
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 localizationCytoplasm > cytosol. Nucleus. Cytosolic under unstressed conditions, translocates into the nucleus upon induction by electrophilic agents.
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Anti-Nrf2 (phospho S40) antibody [EP1809Y] images
Overlay histogram showing Jurkat cells stained with ab76026 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76026, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
All lanes : Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026) at 1/20000 dilution
Lane 1 : HepG2 cell lysate
Lane 2 : HepG2 cell lysate treated with AP
Lysates/proteins at 10 µg per lane.
HRP labelled goat anti-rabbit at 1/1000 dilution
Predicted band size : 68 kDa
Observed band size : 90 kDa (why is the actual band size different from the predicted?)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab76026 at 1/100 dilution.
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.
ICC/IF image of ab76026 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76026, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab76026 showing positive staining in Breast carcinoma tissue.
ab76026 showing positive staining in Cervical carcinoma tissue.
ab76026 showing positive staining in Ovarian carcinoma tissue.
ab76026 showing positive staining in Normal tonsil tissue.
References for Anti-Nrf2 (phospho S40) antibody [EP1809Y] (ab76026)
This product has been referenced in:
- Ji L et al. Nrf2 pathway regulates multidrug-resistance-associated protein 1 in small cell lung cancer. PLoS One 8:e63404 (2013). IHC-P ; Human . Read more (PubMed: 23667609) »
- Inui T et al. Association of p62/SQSTM1 Excess and Oral Carcinogenesis. PLoS One 8:e74398 (2013). WB ; Human . Read more (PubMed: 24086340) »