NRF2, NFE2L2 Recombinant antibody

• Featured Product
• KD/KO Validated

NRF2, NFE2L2 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ChIP, ELISA

Conjugate

Unconjugated

Publications

60

CloneNo.

1I21

Cat no : 80593-1-RR

Synonyms

NRF2, NFE2L2, HEBP1, Nrf 2, NFE2-related factor 2

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Validation Data Gallery

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  WB analysis using 80593-1-RR

  Untreated and MG132 treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80593-1-RR (NRF2, NFE2L2 antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 80593-1-RR

  Untreated and MG132 treated HepG2 cells were subjected to SDS PAGE followed by western blot with 80593-1-RR (NRF2, NFE2L2 antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 80593-1-RR

  Untreated and MG132 treated HeLa cells were subjected to SDS PAGE followed by western blot with 80593-1-RR (NRF2, NFE2L2 antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 80593-1-RR

  Untreated and MG132 treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 80593-1-RR (NRF2, NFE2L2 antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 80593-1-RR

  IP result of anti-NRF2, NFE2L2 (IP:80593-1-RR, 4ug; Detection:80593-1-RR 1:700) with HeLa cells lysate 1520 ug.

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  IHC staining of human ovary tumor using 80593-1-RR

  Immunohistochemical analysis of paraffin-embedded human ovary tumor tissue slide using 80593-1-RR (NRF2, NFE2L2 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human ovary tumor using 80593-1-RR

  Immunohistochemical analysis of paraffin-embedded human ovary tumor tissue slide using 80593-1-RR (NRF2, NFE2L2 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human pancreas cancer using 80593-1-RR

  Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 80593-1-RR (NRF2, NFE2L2 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human pancreas cancer using 80593-1-RR

  Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 80593-1-RR (NRF2, NFE2L2 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 80593-1-RR

  Immunofluorescent analysis of (-20°C Methanol) fixed HepG2 cells using NRF2, NFE2L2 antibody (80593-1-RR, Clone: 1I21 ) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  IF Staining of HepG2 using 80593-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using NRF2, NFE2L2 antibody (80593-1-RR, Clone: 1I21 ) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HepG2 using 80593-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed MG132 treated HepG2 cells using NRF2, NFE2L2 antibody (80593-1-RR, Clone: 1I21 ) at dilution of 1:600 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• 

  FC experiment of MCF-7 using 80593-1-RR

  1X10^6 MCF-7 cells were intracellularly stained with 0.4 ug Anti-Human NRF2, NFE2L2 (80593-1-RR, Clone:1I21) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 80593-1-RR

  Biolayer interferometry (BLl) kinetic assays of 80593-1-RR against Human NRF2, NFE2L2 were performed. The affinity constant is 0.191 nM.

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Tested Applications

• Positive WB detected in: MG132 treated HEK-293 cells, MG132 treated HepG2 cells, MG132 treated HeLa cells, MG132 treated NIH/3T3 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human ovary tumor tissue, human pancreas cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: MG132 treated HepG2 cells, HepG2 cells
• Positive FC (Intra) detected in: MCF-7 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:5000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/ICC : 1:300-1:1200
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 2 publications below
• WB: See 55 publications below
• IHC: See 6 publications below
• IF: See 10 publications below
• ChIP: See 1 publications below

Product Information

80593-1-RR targets NRF2, NFE2L2 in WB, IHC, IF/ICC, FC (Intra), IP, ChIP, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: NRF2, NFE2L2 fusion protein Ag9489
• Full Name: nuclear factor (erythroid-derived 2)-like 2
• Calculated Molecular Weight: 605 aa, 68 kDa
• Observed Molecular Weight: 110 kDa, 68 kDa
• GenBank Accession Number: BC011558
• Gene Symbol: NRF2
• Gene ID (NCBI): 4780
• RRID: AB_2918904
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

NRF2, also named as NFE2L2, belongs to the bZIP family and CNC subfamily. It is a transcription activator that binds to antioxidant response (ARE) elements in the promoter regions of target genes. NRF2 is important for the coordinated up-regulation of genes in response to oxidative stress. It 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. Nrf2 is a key player in the regulation of genes encoding for many antioxidative response enzymes.The expression of NRF2 may be induced under oxidative stress (PMID:14567983).In lung cancer, Nrf2 activation in malignant cells has been associated with tumor progression and chemotherapy resistance(PMID:20534738). Identifying patients with abnormal NRF2 expression may be important for selection for chemotherapy in NSCLC. As new investigators break into the emerging field of Nrf2 research, confusion regarding the correct migratory pattern of Nrf2 is causing doubts about the accuracy and reproducibility of published results. This letter provides solid evidence that the actually observed molecular weight of Nrf2 is about 70kDa and 95-110 kDa. (PMID: 22703241).

Publications

Species	Application	Title
mouse	WB	Cell Host Microbe; Liberation of daidzein by gut microbial β-galactosidase suppresses acetaminophen-induced hepatotoxicity in mice; Authors - Yunong Zeng
human	WB	J Med Virol; SARS-CoV-2 nucleocapsid protein enhances the level of mitochondrial reactive oxygen species; Authors - Haiyun Yu
mouse	WB	Sci Total Environ; Effects of chronic low-level lead (Pb) exposure on cognitive function and hippocampal neuronal ferroptosis: An integrative approach using bioinformatics analysis, machine learning, and experimental validation; Authors - Yingsi Cao
human,mouse	WB	Br J Pharmacol; Leonurine hydrochloride alleviates ferroptosis in cisplatin-induced acute kidney injury by activating the Nrf2 signaling pathway.; Authors - Jianqiang Hu ; KO Validated
human	WB	Antioxidants (Basel); Glycolysis Aids in Human Lens Epithelial Cells' Adaptation to Hypoxia; Authors - Yuxin Huang
human	WB	Mol Med; β-aminoisobutyrics acid, a metabolite of BCAA, activates the AMPK/Nrf-2 pathway to prevent ferroptosis and ameliorates lung ischemia-reperfusion injury; Authors - Ziyue Zhang