Nrf2 Antibody | Affinity Biosciences

Product:	Nrf2 Antibody
Catalog:	AF7006
Description:	Rabbit polyclonal antibody to Nrf2
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.:	68kDa,125kDa; 68kD(Calculated).
Uniprot:	Q16236
RRID:	AB_2835314

View similar products>>

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
50ul	$250	In stock
100ul	$350	In stock
200ul	$450	In stock

Lead Time: Same day delivery

For pricing and ordering contact:
Local distributors

Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, 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(90%), Horse(80%), Sheep(80%), Rabbit(100%), Dog(90%)

Clonality:

Polyclonal

Specificity:

Nrf2 Antibody detects endogenous levels of total Nrf2.

RRID:

AB_2835314
Cite Format: Affinity Biosciences Cat# AF7006, RRID:AB_2835314.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

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.

Description:

NRF2 a transcription factor that regulates basal expression and antioxidant induction of a transcription factor for NAD(P)H:quinone oxidoreductase-1 (NQO1) and other detoxifying genes. Targeted for proteasomal degradation by INrf2.

Sequence:

Q16236 NF2L2_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

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

Rabbit

100

Pig

100

Bovine

Dog

Horse

Sheep

Xenopus

Chicken

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q16236 As Substrate

Q16236

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

Research Backgrounds

Q16236_NF2L2_HUMAN

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).

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.

References

1). Sulforaphane elicts dual therapeutic effects on Renal Inflammatory Injury and crystal deposition in Calcium Oxalate Nephrocalcinosis. Theranostics (PubMed: 32641994) [IF=12.4]

Application: IHC Species: mice Sample: tubular epithelial cells

Figure 2. Nrf2 significantly suppresses TLR4 and IRF1 levels in a mouse model of CaOx nephrocalcinosis. (A) IHC staining of kidney Nrf2, TLR4, and IRF1 in CaOx nephrocalcinosis mice treated with SFN (400× magnification; scale bar: 40 µm). SFN treatment increases Nrf2 and inhibits the expression of TLR4 and IRF1 in CaOx nephrocalcinosis mouse tubular epithelial cells. (B) Quantification of IHC staining of Nrf2, TLR4, and IRF1 in SFN treated CaOx nephrocalcinosis mouse model. (C) qPCR detection of Nrf2, TLR4, and IRF1 expression in SFN-treated CaOx nephrocalcinosis mouse kidney samples and comparison with normal controls. (D, E) Pearson’s correlation analysis of Nrf2 levels relative to TLR4 and IRF1. Data represent the mean ± standard error (SE) of three independent experiments. *P < 0.05; **P < 0.01, as determined by one-way ANOVA (B, C) or Pearson’s correlation (D, E).

Application: WB Species: mice Sample: BMDMs

Figure 3. Nrf2 suppresses TLR4 and IRF1 levels and promotes M2Mϕ polarization in vitro. (A) BMDM and COM-TECs coculture schematic diagram. Western blot (B) and qPCR (C) analyses of Nrf2, TLR4, IRF1, iNOS, and ARG-1 levels in BMDMs co-cultured with increasing COM dose stimulated TECs. GAPDH was used as an internal control. (D, F) Western blot detection of Nrf2, TLR4, IRF1, iNOS, and ARG-1 levels following SFN treatment or Nrf2 upregulation/downregulation in BMDMs co-cultured with COM-stimulated TECs. GAPDH was used as an internal control. (E, G) The distribution of iNOS (green) and ARG-1 (red) in BMDMs according to immunofluorescence. (H, I) Flow cytometric analysis of the polarization state of BMDMs using anti-CD11c and anti-CD206 in F4/80+ and CD11b+ cells. The data are shown as the mean ± standard error (SE) of three independent experiments. *P < 0.05; **P < 0.01, as determined by Student’s t test (C) or one-way ANOVA (E, G, H, I).

2). Cytotoxicity of adducts formed between quercetin and methylglyoxal in PC-12 cells. Food Chemistry (PubMed: 33706136) [IF=8.8]

Application: WB Species: Rat Sample: PC-12 cells

Fig. 5. Effect of treatments of MGO, Que-mono-MGO, and Que-di-MGO on the expression levels of apoptotic markers and components of AKT and Nrf2-HO-1/NQO-1 signaling pathways. Significant differences (p < 0.05) between samples of different treatments are marked with different letters on each column.

3). Naringenin protects against iron overload-induced osteoarthritis by suppressing oxidative stress. PHYTOMEDICINE (PubMed: 35905566) [IF=7.9]

4). The effect of monotropein on alleviating cisplatin-induced acute kidney injury by inhibiting oxidative damage, inflammation and apoptosis. BIOMEDICINE & PHARMACOTHERAPY (PubMed: 32574971) [IF=7.5]

Application: WB Species: Human Sample: kidney tissue

Fig. 2. Pretreatment with monotropein protected against renal oxidative stress induced by cisplatin. The level of serum GSH(A). The level of serum MDA(B). The level of serum SOD(C). The level of serum CAT(D). The protein expressions of Nrf2, HO-1 and NQO1 (E) were detected by Western blot in kidney tissue. The protein expressions of Nrf2 (F), HO-1 (G) and NQO1 (H) were quantitated by Image software. Data are showed as mean ± S.E.M of 6 mice in each group. #P < 0.05, ##P < 0.01 vs. Normal group; *P < 0.05, **P < 0.01 vs. cisplatin group.

5). Human umbilical cord mesenchymal stem cells (hUC-MSCs) alleviate paclitaxel-induced spermatogenesis defects and maintain male fertility. Biological Research (PubMed: 37574561) [IF=6.7]

Application: IHC Species: Mouse Sample:

Fig. 9 Expressions of NRF2, SIRT1 and antioxidant molecules of CAT, SOD1, PRDX6 in control, PTX treatment and PTX + hUC-MSCs treatment mice testis. The mice were treated with PTX or hUC-MSCs, and samples were collected 2 weeks later. A: Testicular expression of NRF2 and SIRT1 in control, PTX and PTX + hUC-MSCs treated mice testes; B: Expressions of CAT, SOD1, and PRDX6 in mice testes of control, PTX and PTX + hUC-MSCs group were detected by Western blotting, and analyzed by One-Way ANOVA; C: Gene expressions of Cat, Sod1, and Prdx6; p value less than 0.05 was considered significance;

6). Metformin suppresses oxidative stress induced by high glucose via activation of the Nrf2/HO-1 signaling pathway in type 2 diabetic osteoporosis. LIFE SCIENCES (PubMed: 36279968) [IF=6.1]

7). Hepatoprotective and Anti-Oxidative Effects of Total Flavonoids From Qu Zhi Qiao (Fruit of Citrus Paradisi cv.Changshanhuyou) on Nonalcoholic Steatohepatitis In Vivo and In Vitro Through Nrf2-ARE Signaling Pathway. Frontiers in Pharmacology (PubMed: 32390839) [IF=5.6]

Application: WB Species: mouse Sample: liver

FIGURE 5 | Expression of the targeted proteins in mice liver. Values are shown as mean ± SD (n = 8). Data (mean ± SD) with different case letters are statistically different with each other at LSD multiple comparisons, which with red letters differed significantly as compared to model group level. uppercase letter p < 0.01 vs.another group.

Application: IHC Species: mouse Sample: liver

FIGURE 4 | Immunohistochemistry observation of nuclear factor erythroid 2-related factor 2 (Nrf2) expressions in livers of mice. Images were obtained at 400×magnification (scale bar=50mm).

8). Dehydrocostus Lactone Suppresses Dextran Sulfate Sodium-Induced Colitis by Targeting the IKKα/β-NF-κB and Keap1-Nrf2 Signalling Pathways. Frontiers in Pharmacology (PubMed: 35321327) [IF=5.6]

Application: WB Species: mouse Sample: RAW264.7 macrophages

FIGURE 6 | DCL enhances the activation of Keap1/Nrf2/HO-1 in LPS/IFNγ-stimulated RAW264.7 macrophages. RAW264.7 cells were treated with DCL (1, 3, and 9 μM) or curcumin (5 μM) for 1 h, and followed by LPS/IFNγ incubation for additional 8 h. The protein expression of Keap1 (A), Nrf2 (C), and HO-1 (G) were measured by western blot.

9). Nepeta angustifolia attenuates responses to vascular inflammation in high glucose-induced human umbilical vein endothelial cells through heme oxygenase-1 induction. JOURNAL OF ETHNOPHARMACOLOGY (PubMed: 30419276) [IF=5.4]

10). Inhibition of micro RNA miR-122-5p prevents lipopolysaccharide-induced myocardial injury by inhibiting oxidative stress, inflammation and apoptosis via targeting GIT1. Bioengineered (PubMed: 34002676) [IF=4.9]

Application: WB Species: Rat Sample: heart tissues

Figure 4. Inhibition of micro RNA miR-122-5p suppresses lipopolysaccharide (LPS)-induced oxidative stress and inflammatory response. (a) Reactive oxygen species (ROS) production was determined by a flow cytometer. (b) The enzymes catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-px) activities were measured using the manufacturer’s kits. (c) Western blot was performed to measure the nuclear factor erythroid 2-related factor 2 (Nrf-2) expression in the cytoplasm and nucleus. (d) Real-time quantitative PCR (RT-qPCR) was used for heme oxygenase-1 (HO-1) and NAD(p)H: quinone oxidoreductase 1 (NQO-1) levels. (e) The concentrations of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and IL-1β were quantified by enzyme-linked immunosorbent assay (ELISA) kits. ***p < 0.001 versus control; +p < 0.05, ++p < 0.01, +++p < 0.001 versus LPS + NC inhibitor

Restrictive clause

Affinity Biosciences tests all products strictly. Citations are provided as a resource for additional applications that have not been validated by Affinity Biosciences. Please choose the appropriate format for each application and consult Materials and Methods sections for additional details about the use of any product in these publications.

For Research Use Only.
Not for use in diagnostic or therapeutic procedures. Not for resale. Not for distribution without written consent. Affinity Biosciences will not be held responsible for patent infringement or other violations that may occur with the use of our products. Affinity Biosciences, Affinity Biosciences Logo and all other trademarks are the property of Affinity Biosciences LTD.

Nrf2 Antibody - #AF7006