Product Name
Phospho-Nrf2 (S40) Recombinant Rabbit Monoclonal Antibody [SU0334] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Ser40 of Human Nrf2 aa 20-69 / 605.
Species Reactivity
Human (Predicted: Mouse, Rat)
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 68 kDa
Positive Control
HepG2 cell lysate, Raji cell lysate, HepG2, Hela, A549, human breast carcinoma tissue, human tonsil tissue, human kidney tissue, K562.
Storage Instructions
Store at 2-8℃. Avoid freeze.
Purification Method
Protein A affinity purified.
Function
Nuclear factor erythroid 2-related factor 2 (NRF2), also known as nuclear factor erythroid-derived 2-like 2, is a transcription factor that in humans is encoded by the NFE2L2 gene. NRF2 is a basic leucine zipper (bZIP) protein that may regulate the expression of antioxidant proteins that protect against oxidative damage triggered by injury and inflammation, according to preliminary research.[6] In vitro, NRF2 binds to antioxidant response elements (AREs) in the promoter regions of genes encoding cytoprotective proteins. NRF2 induces the expression of heme oxygenase 1 in vitro leading to an increase in phase II enzymes. NRF2 also inhibits the NLRP3 inflammasome. NRF2 appears to participate in a complex regulatory network and performs a pleiotropic role in the regulation of metabolism, inflammation, autophagy, proteostasis, mitochondrial physiology, and immune responses.
Background References
1. He F et al. NRF2, a Transcription Factor for Stress Response and Beyond. Int J Mol Sci. 2020 Jul
2. Ulasov AV et al. Nrf2/Keap1/ARE signaling: Towards specific regulation. Life Sci. 2022 Feb
Sequence Similarity
Belongs to the bZIP family. CNC subfamily.
Tissue Specificity
Widely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.
Post-translational Modification
Ubiquitinated in the cytoplasm by the BCR(KEAP1) E3 ubiquitin ligase complex leading to its degradation. In response to oxidative stress, electrophile metabolites, such as sulforaphane, modify KEAP1, leading to inhibit activity of the BCR(KEAP1) complex, promoting NFE2L2/NRF2 nuclear accumulation and activity. In response to autophagy, the BCR(KEAP1) complex is inactivated (By similarity).; Phosphorylation of Ser-40 by PKC in response to oxidative stress dissociates NFE2L2 from its cytoplasmic inhibitor KEAP1, promoting its translocation into the nucleus.; Acetylation at Lys-596 and Lys-599 increases nuclear localization whereas deacetylation by SIRT1 enhances cytoplasmic presence.; Glycation impairs transcription factor activity by preventing heterodimerization with small Maf proteins. Deglycation by FN3K restores activity.
Subcellular Location
Cytoplasm, cytosol, Nucleus.
Synonyms
erythroid derived 2 antibody
HEBP1 antibody
like 2 antibody
NF E2 related factor 2 antibody
NF-E2-related factor 2 antibody
NF2L2_HUMAN antibody
NFE2 related factor 2 antibody
NFE2-related factor 2 antibody
Nfe2l2 antibody
Nrf 2 antibody
Expand
erythroid derived 2 antibody
HEBP1 antibody
like 2 antibody
NF E2 related factor 2 antibody
NF-E2-related factor 2 antibody
NF2L2_HUMAN antibody
NFE2 related factor 2 antibody
NFE2-related factor 2 antibody
Nfe2l2 antibody
Nrf 2 antibody
NRF2 antibody
Nuclear factor (erythroid derived 2) like 2 antibody
Nuclear factor antibody
nuclear factor erythroid 2 like 2 antibody
Nuclear factor erythroid 2 related factor 2 antibody
Nuclear factor erythroid 2-related factor 2 antibody
Nuclear factor erythroid derived 2 like 2 antibody
Collapse
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This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Western blot analysis of Phospho-Nrf2 (S40) on different lysates with Rabbit anti-Phospho-Nrf2 (S40) antibody (ET1608-28) at 1/2,000 dilution.
Lane 1: HepG2 cell lysate
Lane 2: Raji cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 68 kDa
Observed band size: 100 kDa
Exposure time: 30 seconds;
8% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-28) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HepG2 cells labeling Phospho-Nrf2 (S40) with Rabbit anti-Phospho-Nrf2 (S40) antibody (ET1608-28) at 1/250 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-Nrf2 (S40) antibody (ET1608-28) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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☑ Cell treatment (CT),Relative expression (RE)
This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-Nrf2 (S40) antibody [SU0334] (ET1608-28).
The section was pre-treated using heat mediated antigen retrieval with 10 mM sodium citrate buffer (pH 6.0). The section were untreated (A, C) and treated (B) with λ-PPase at 1/25 dilution at 30℃ for 30 minutes after antigen retrieval. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28, 1/200) for 30 minutes at room temperature. Goat anti-Rabbit IgG-HRP UltraPolymer antibody (HA1119) was used for 20 minutes at room temperature. DAB was used as the chromogen. The section were counterstained with hematoxylin and mounted with DPX. PBS was used instead of the primary antibody as the negative control, and is shown in the inset (C).
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This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Phospho-Nrf2 (S40) antibody (ET1608-28) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Phospho-Nrf2 (S40) antibody (ET1608-28) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Phospho-Nrf2 (S40) antibody (ET1608-28) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Phospho-Nrf2 (S40) antibody (ET1608-28) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-Phospho-Nrf2 (S40) antibody (ET1608-28) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1608-28, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of Phospho-Nrf2 (S40) was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-28, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
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