Phospho-Nrf2 (Ser40) Antibody - #DF7519

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Product: Phospho-Nrf2 (Ser40) Antibody
Catalog: DF7519
Description: Rabbit polyclonal antibody to Phospho-Nrf2 (Ser40)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Rabbit, Dog, Chicken
Mol.Wt.: 90 kDa; 68kD(Calculated).
Uniprot: Q16236
RRID: AB_2841018

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:3000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(91%), Horse(89%), Rabbit(82%), Dog(100%), Chicken(91%)
Clonality:
Polyclonal
Specificity:
Phospho-Nrf2 (Ser40) Antibody detects endogenous levels of Nrf2 only when phosphorylated at S40.
RRID:
AB_2841018
Cite Format: Affinity Biosciences Cat# DF7519, RRID:AB_2841018.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

erythroid derived 2; HEBP1; like 2; NF E2 related factor 2; NF-E2-related factor 2; NF2L2_HUMAN; NFE2 related factor 2; NFE2-related factor 2; Nfe2l2; Nrf 2; NRF2; Nuclear factor (erythroid derived 2) like 2; Nuclear factor; nuclear factor erythroid 2 like 2; Nuclear factor erythroid 2 related factor 2; Nuclear factor erythroid 2-related factor 2; Nuclear factor erythroid derived 2 like 2;

Immunogens

Immunogen:
Uniprot:

Q16236(NF2L2_HUMAN) >>Visit HPA database.

Gene(ID):
NFE2L2(4780)
Expression:
Q16236 NF2L2_HUMAN:

Widely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.

Sequence:
MMDLELPPPGLPSQQDMDLIDILWRQDIDLGVSREVFDFSQRRKEYELEKQKKLEKERQEQLQKEQEKAFFAQLQLDEETGEFLPIQPAQHIQSETSGSANYSQVAHIPKSDALYFDDCMQLLAQTFPFVDDNEVSSATFQSLVPDIPGHIESPVFIATNQAQSPETSVAQVAPVDLDGMQQDIEQVWEELLSIPELQCLNIENDKLVETTMVPSPEAKLTEVDNYHFYSSIPSMEKEVGNCSPHFLNAFEDSFSSILSTEDPNQLTVNSLNSDATVNTDFGDEFYSAFIAEPSISNSMPSPATLSHSLSELLNGPIDVSDLSLCKAFNQNHPESTAEFNDSDSGISLNTSPSVASPEHSVESSSYGDTLLGLSDSEVEELDSAPGSVKQNGPKTPVHSSGDMVQPLSPSQGQSTHVHDAQCENTPEKELPVSPGHRKTPFTKDKHSSRLEAHLTRDELRAKALHIPFPVEKIINLPVVDFNEMMSKEQFNEAQLALIRDIRRRGKNKVAAQNCRKRKLENIVELEQDLDHLKDEKEKLLKEKGENDKSLHLLKKQLSTLYLEVFSMLRDEDGKPYSPSEYSLQQTRDGNVFLVPKSKKPDVKKN

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Dog
100
Bovine
91
Chicken
91
Horse
89
Rabbit
82
Xenopus
73
Sheep
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q16236 As Substrate

Site PTM Type Enzyme
S40 Phosphorylation P05771 (PRKCB) , P17252 (PRKCA)
K44 Ubiquitination
K64 Ubiquitination
T80 Phosphorylation
S215 Phosphorylation
S301 Phosphorylation
S344 Phosphorylation P49841 (GSK3B)
S347 Phosphorylation P49841 (GSK3B)
S351 Phosphorylation
S356 Phosphorylation
T395 Phosphorylation Q00535 (CDK5)
S408 Phosphorylation
S433 Phosphorylation Q00535 (CDK5)
R437 Methylation
K438 Acetylation
T439 Phosphorylation Q00535 (CDK5)
K443 Acetylation
K445 Acetylation
S447 Phosphorylation
K462 Acetylation
K462 Ubiquitination
K472 Acetylation
K487 Acetylation
K506 Acetylation
K508 Acetylation
K516 Acetylation
K516 Ubiquitination
K518 Acetylation
K518 Sumoylation
K518 Ubiquitination
K533 Acetylation
K536 Acetylation
K538 Acetylation
K541 Acetylation
K543 Acetylation
K548 Acetylation
K548 Ubiquitination
K554 Acetylation
K554 Ubiquitination
K555 Acetylation
S558 Phosphorylation
T559 Phosphorylation
Y561 Phosphorylation
K574 Ubiquitination
Y576 Phosphorylation P06241 (FYN)
S577 Phosphorylation
K596 Acetylation
K599 Acetylation

Research Backgrounds

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. In normal conditions, ubiquitinated and degraded in the cytoplasm by the BCR(KEAP1) complex. 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. 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. 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.

PTMs:

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.
Note: Cytosolic under unstressed conditions: ubiquitinated and degraded by the BCR(KEAP1) E3 ubiquitin ligase complex (PubMed:15601839, PubMed:21196497). Translocates into the nucleus upon induction by electrophilic agents that inactivate the BCR(KEAP1) E3 ubiquitin ligase complex (PubMed:21196497).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Widely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.

Subunit Structure:

Heterodimer; heterodimerizes with small Maf proteins (By similarity). Interacts (via the bZIP domain) with MAFG and MAFK; required for binding to antioxidant response elements (AREs) on DNA (By similarity). Interacts with KEAP1; the interaction is direct and promotes ubiquitination by the BCR(KEAP1) E3 ubiquitin ligase complex. Forms a ternary complex with PGAM5 and KEAP1. Interacts with EEF1D at heat shock promoter elements (HSE). Interacts via its leucine-zipper domain with the coiled-coil domain of PMF1. Interacts with CHD6; involved in activation of the transcription (By similarity). Interacts with ESRRB; represses NFE2L2 transcriptional activity (By similarity).

(Microbial infection) Interacts with herpes virus 8 protein LANA1.

Family&Domains:

The ETGE motif, and to a lower extent the DLG motif, mediate interaction with KEAP1.

Belongs to the bZIP family. CNC subfamily.

Research Fields

· Genetic Information Processing > Folding, sorting and degradation > Protein processing in endoplasmic reticulum.   (View pathway)

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.   (View pathway)

References

1). Amelioratory effects of astragaloside IV on hepatocarcinogenesis via Nrf2-mediated pSmad3C/3L transformation. Phytomedicine (PubMed: 37301185) [IF=7.9]
2). Phosphodiesterase 4B is required for NLRP3 inflammasome activation by positive feedback with Nrf2 in the early phase of LPS- induced acute lung injury. Free Radical Biology and Medicine (PubMed: 34644617) [IF=7.4]
3). Eriodictyol ameliorates cognitive dysfunction in APP/PS1 mice by inhibiting ferroptosis via vitamin D receptor-mediated Nrf2 activation. Molecular Medicine (PubMed: 35093024) [IF=5.7]

Application: WB    Species: Mouse    Sample: brain

Fig. 7 Eriodictyol up-regulates VDR expression and activates the Nrf2/HO-1 signaling pathway. A VDR expression in the mouse brain was detected using immunohistochemical staining. Then, B the VDR protein expression level in the mouse cortex and hippocampus was measured using Western blotting. C The VDR protein expression level in HT-22 cells was measured using Western blotting, and D its relative mRNA expression level was detected using q-PCR. E The levels of Nrf2, p-Nrf2 and HO-1 in the mouse cortex and hippocampus were measured using Western blotting. F The levels of Nrf2, p-Nrf2 and HO-1 in HT-22 cells were measured using Western blotting. G The level of Nrf2 in the nucleus and cytoplasm of HT-22 cells was detected using Western blotting, and Lamin B1 and β-actin were used as loading controls for the nuclear and cytoplasmic proteins, respectively. The data are presented as the means ± SD of three experiments. **P < 0.01 and ***P < 0.001 compared with the Aβ1–42 oligomer group. ##P < 0.01 compared with the control group
4). The protective role of IL-1Ra on intestinal ischemia reperfusion injury by anti-oxidative stress via Nrf2/HO-1 pathway in rat. Biomedical Journal (PubMed: 30987703) [IF=5.5]

Application: WB    Species: rat    Sample: intestine

Fig. 6 | IL-1Ra activates Nrf2/HO-1 signaling in I/R intestine. (A) The protein expression levels of p-Nrf2, Nrf2 and HO-1.
5). Astragaloside IV inhibits hepatocellular carcinoma by continually suppressing the development of fibrosis and regulating pSmad3C/3L and Nrf2/HO-1 pathways. JOURNAL OF ETHNOPHARMACOLOGY (PubMed: 34157326) [IF=5.4]

Application: WB    Species: Mice    Sample: liver tissue

Fig. 5. AS-IV modulates pSmad3C/3L and Nrf2/HO-1 pathways in mice treated with DCC for 12 weeks. (A), (B), (C), (F), (G), (H), (I), (M) Hepatic expression and distribution pattern of proteins (TGF-β1, pSmad3C, pSmad3L, pNrf2) by immunofluorescent staining, and relative fluorescent intensity of TGF-β1, pSmad3C, pSmad3L, pNrf2 in the liver (Magnification × 400); (D), (E), (K), (L) Representative immunoblots and relative protein levels of TGF-β1, pSmad3C, pSmad3L, pSmad2C, pSmad2L, PAI-1, pNrf-2, Nrf-2, HO-1, NQO1. (J), (N) The Nrf-2 expression was evaluated by immunohistochemistry (Magnification × 400), and quantitation (% of the positively stained area) of Nrf2 by immunohistochemistry; (O) the correlation analysis of pSmad3C and pNrf2; (P) the correlation analysis of pSmad3L and pNrf2. Data are presented as mean ± SD (n = 3 for each group). #P < 0.05 VS. Control. # #P < 0.01 VS. Control. *P < 0.05 VS. Model. **P < 0.01 VS. Model.
6). Zhen-Wu decoction and lactiflorin, an ingredient predicted by in silico modelling, alleviate uremia induced cardiac endothelial injury via Nrf2 activation. JOURNAL OF ETHNOPHARMACOLOGY (PubMed: 35963415) [IF=5.4]
7). A diarylheptanoid compound from Alpinia officinarum Hance ameliorates high glucose-induced insulin resistance by regulating PI3K/AKT-Nrf2-GSK3β signaling pathways in HepG2 cells. JOURNAL OF ETHNOPHARMACOLOGY (PubMed: 35605918) [IF=5.4]
8). Andrographolide Inhibits Static Mechanical Pressure-Induced Intervertebral Disc Degeneration via the MAPK/Nrf2/HO-1 Pathway. Drug design, development and therapy (PubMed: 36845666) [IF=4.8]
9). Therapeutic potential of luteolin on impaired wound healing in streptozotocin-induced rats. Biomedicines (PubMed: 34209369) [IF=4.7]

Application: WB    Species: Rat    Sample: skin tissue

Figure 6. Effects of luteolin on protein expression levels of oxidative enzymes of the skin wound on day 14. ( A ) Tissues were homogenized in RIPA buffer, subjected to immunoblotting with anti- Nrf2, anti-phospho-Nrf2, anti-SOD1, and anti-GSH-Px antibodies. ( B ) Quantitative results of Nrf2, phospho-Nrf2, SOD1, and GSH-Px protein levels were adjusted to β -actin protein level. Values of individual groups are presented as mean ± SEM. * p < 0.05, compared with the NDM group. # p < 0.05, compared with the DM group.
10). Mechanism for antiParkinsonian effect of resveratrol: Involvement of transporters, synaptic proteins, dendrite arborization, biochemical alterations, ER stress and apoptosis. FOOD AND CHEMICAL TOXICOLOGY (PubMed: 34302886) [IF=4.3]

Application: WB    Species: Rat    Sample: brain tissue

Fig. 13. Western blot results for the detection of EGF receptor, HIF1A and NOS3. Sham versus I/R, # P < 0.05; HSYA versus I/R, **P < 0.01; a–b EGF receptor; c–d HIF1A; e–f NOS3.
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