Invitrogen

Nrf2 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

1 Published Figure

4 References

View all (22) Nrf2 antibodies

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Questions & Answers

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Nrf2 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of NRF2 (green) HEK293T cells. Cells fixed with 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with an NRF2 polyclonal antibody (Product # PA5-27882) at a dilution of 1:100 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with a fluorophore-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification. Data courtesy of the Innovators Program.

Nrf2 Antibody in Immunocytochemistry (ICC) — Immunofluorescence analysis of Nuclear factor erythroid 2-related factor 2 was performed using 70% confluent log phase Hep G2 cells treated with TBHQ (100 micromolar for 4h). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Nrf2 Polyclonal Antibody (Product # PA5-27882) at 1:100 dilution in 0.1% BSA, incubated at 4-degree Celsius overnight, and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing nuclear translocation of NRF2 upon TBHQ treatment. Panel e represents untreated cells showing predominant cytoplasmic localization. Panel f represents control cells with no primary antibody to assess the background. The images were captured at 60X magnification.

Nrf2 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of Nrf2 was performed in paraffin-embedded human breast carcinoma tissue using Nrf2 Polyclonal Antibody (Product # PA5-27882) at a dilution of 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Nrf2 Antibody in Western Blot (WB) — Western blot was performed using Anti-Nrf2 Polyclonal Antibody (Product # PA5-27882) and a 80kDa band corresponding to Nuclear factor erythroid 2-related factor 2 was observed to be upregulated upon various treatments across the cell lines tested. Whole-cell extracts (30 µg lysate) of U-2 OS (Lane 1), U-2 OS treated with MG-132 (10 micromolar for 10h) (Lane 2), MDA-MB-231 (Lane 3), MDA-MB-231 treated with TBHQ (30 micromolar for 4h) (Lane 4), Hep G2 (Lane 5), Hep G2 treated with TBHQ (100 micromolar for 4h) (Lane 6), HEK-293 (Lane 7), HEK-293 treated with TBHQ (25 micromolar for 4h) (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Nrf2 Antibody in Immunoprecipitation (IP) — Immunoprecipitation of NRF2 in HepG2 whole cell lysates using 5 µg of NRF2 polyclonal antibody (Product # PA5-27882). Western blot analysis performed using NRF2 polyclonal antibody (Product # PA5-27882).

Nrf2 Antibody in ChIP assay (ChIP) — ChIP assay analysis of Nrf2 was performed in HepG2 chromatin extracts using 5 µg of either normal rabbit IgG or Nrf2 Polyclonal Antibody (Product # PA5-27882). The precipitated DNA was detected by PCR with primer set targeting to GCLC gene locus.

Nrf2 Antibody in Cell treatment — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using Nrf2 Polyclonal Antibody (Product # PA5-27882), shows upregulation of its expression upon MG-132 and TBHQ treatments across the cell lines tested. {TM}

Nrf2 Antibody in Knockdown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. U-2 OS cells were transfected with Nuclear factor erythroid 2-related factor 2 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-Nrf2 Polyclonal Antibody (Product # PA5-27882). {KD}

Nrf2 Antibody — Fig. 5 Loss of GSH content in microglia influences oxidative stress but not cell viability. EOC13.31 microglia cells were divided into six groups: either untreated (control) or pretreated with 500 muM NAC or 200 muM BSO for 18 h, prior to 22 mM EtOH exposure for 6 h. Confocal images (20x) of cells labeled with CellROX Deep Red Reagent to detect ROS generation ( a ). Semi-quantification analysis of ROS fluorescence intensity ( b ). Intracellular GSH levels were determined fluorometrically by labeling cells with 50 muM Monochlorobimane ( c ). Western blot analysis for Nrf-2 expression and nuclear translocation, where Lamin B1 and beta actin was used as a loading control for nuclear and cytosolic fraction, respectively ( d ). Effect of the above treatment regimen on cell viability (MTS assay) ( e ). Values are presented as mean average +- SEM three independent experiments. Student-Newman-Keuls comparison analysis was used (* p <= 0.05; ** p <= 0.005)

Product Details

Applications

Tested Dilution

Publications

ChIP assay (ChIP)

5 µg

Immunocytochemistry (ICC)

1:100-1:1000

Immunofluorescence (IF)

1:100-1:1000

Immunohistochemistry (IHC)

Assay Dependent

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:1000

Immunoprecipitation (IP)

1:100-1:500

Western Blot (WB)

1:500-1:3000

View 3 publications 3 publications

Miscellaneous PubMed (Misc)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Avian, Human, Mouse, Plant, Reptile, Rat, Zebrafish

Published species

Human, Zebrafish

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Recombinant fragment corresponding to a region within amino acids 108 and 413 of Human NRF2

Conjugate

Unconjugated

Form

Liquid

Concentration

0.49 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7, with 20% glycerol, 1% BSA

Contains

0.025% ProClin 300

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2545358

Product Specific Information

Recommended positive controls: C8D30, 3xFlag-human NFE2L2-transfected 293T, MDA-MB-231 nuclear extract, MDA-MB-231 nuclear extract (30 µM tBHQ, 4hr), HepG2 (10 µM MG132 treatment for 15 hr).

Predicted reactivity: Rhesus Monkey (99%), Bovine (84%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

Nfr1 (also designated TCF11), Nrf2 and Nfr3 are bZIP transcription factors that heterodimer with Maf proteins to bind MARE (Maf recognition elements) sequences. Nrf2 is widely expressed and is thought to translocate to the nucleus after treatment with xenobiotics and antioxidants, which stimulate its release from a repressor protein Keap1.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: E4TF1-60; E4TF1A; HEBP1; NF-E2-related factor 2; NFE2-related factor 2; NFT2; nrf2; NRF2A; nuclear factor erythroid 2-like 2; Nuclear factor erythroid 2-related factor 2; nuclear factor erythroid-derived 2-like 2; nuclear factor, erythroid 2-like 2; Nuclear factor, erythroid derived 2, like 2

Gene Aliases: AI194320; HEBP1; NFE2L2; NRF2

UniProt ID: (Human) Q16236, (Mouse) Q60795, (Rat) O54968

Entrez Gene ID: (Human) 4780, (Mouse) 18024, (Rat) 83619

Molecular Function: actin family cytoskeletal protein cytoskeletal protein hydrolase non-motor actin binding protein nucleic acid binding protease serine protease transcription factor