Invitrogen

Nrf2 Polyclonal Antibody

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This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

11 Published Figures

17 References

View all (31) Nrf2 antibodies

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Nrf2 Antibody in Immunocytochemistry (ICC/IF) — 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)) — Nrf2 Polyclonal Antibody detects NRF2 protein at cytoplasm and nucleus by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. NRF2 stained by Nrf2 Polyclonal Antibody (Product # PA5-27882) diluted at 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 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 Western Blot, Immunohistochemistry (WB, IHC) — Fig. 5 TRIM15 stabilizes Nrf2 through binding with Keap1. A HEK293 cells transfected with Flag-TRIM15, HA-Keap1, and Myc-Nrf2 were subjected to immunoprecipitation with HA antibody. Lysates were analyzed by western blotting. B TRIM15 reduced the interaction between Nrf2 and Keap1. Cell lysates were immunoprecipitated with an anti-Keap1 antibody and blotted with an anti-Nrf2 antibody. C - F Subcellular fractionation was used to isolate cytoplasmic and nuclear proteins, and immunoblotting was performed to examine the localization of Nrf2 following the downregulation or overexpression of TRIM15. Nuclear and cytoplasmic levels of Nrf2 are quantified. G Effect of TRIM15 knockdown (H1299 cells) or overexpression (H1650 cells) on the mRNA expression of the Nrf2-regulated genes. NAD(P)H quinone dehydrogenase1(NOQ1), thioredoxin (TXN), peroxiredoxin 1(PRDX1), hemoxygenase 1(HMOX1), glutamate-cysteine ligase catalytic subunit (GCLC), glutathione S-transferase mu1(GSTM1), glutathione S-transferase mu3(GSTM3), ferritin light chain (FTL). H Representative IHC staining images of Nrf2 in the same set of NSCLC tissue slices. Correlation analysis of TRIM15 and Nrf2 expression in NSCLC samples. Spearman correlation coefficients are shown. Scale bars, 100 mum. Statistical analyses were performed by two-tailed unpaired Student's t -test. ** P < 0.01

Nrf2 Antibody in Western Blot, Immunoprecipitation (WB, IP) — Figure 3 SLD-LPS-mediated Keap1 accumulation is correlated with a pro-inflammatory monocyte phenotype. BMDMs were treated with either PBS or SLD-LPS (100 pg/mL) for 5 days. (A) Representative Co-IP result with no lysate IP (just antibody and beads) (Con.) and the whole cell lysate input (WCL) controls. Keap1 antibody was used to pull-down Nrf2 (upper blot), and Nrf2 was used to pull down Keap1 (lower blot). See Supplemental Figures S3 , S4 for un-altered blots. (B-D) The levels of Ly6C (B) , Keap-1 (C) , and CD200R (D) were analyzed and quantified using geometric mean fluorescence intensity (MFI) by flow cytometry. (E-G) Representative western blots of IKKbeta (E) , p-p65 (S536) (F) , and total NF-kappaB p65 (G) in monocytes with quantification on the left. (H) p-p65/total p65 ratio. All data are representative of at least 3 separate experiments ( n = 3, except for IKK-beta where n = 5). Unpaired student t -test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Nrf2 Antibody — 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}

Product Details

PA5-27882

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	View 11 publications 11 publications
Immunohistochemistry (IHC)	-	View 2 publications 2 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	View 2 publications 2 publications
Immunoprecipitation (IP)	1:100-1:500	View 1 publication 1 publication
ChIP assay (ChIP)	5 µg	-
Miscellaneous PubMed (Misc)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Avian, Human, Mouse, Plant, Reptile, Rat, Zebrafish
Published species	Human, Mouse, Pig, 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 View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.41 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.

Product References

E3 ligase TRIM15 facilitates non-small cell lung cancer progression through mediating Keap1-Nrf2 signaling pathway.

Cell communication and signaling : CCS

Liang M,Wang L,Sun Z,Chen X,Wang H,Qin L,Zhao W,Geng B

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Western Blot

Mon May 09 00:00:00 EDT 2022

E3 ligase TRIM15 facilitates non-small cell lung cancer progression through mediating Keap1-Nrf2 signaling pathway.

Cell communication and signaling : CCS

Liang M,Wang L,Sun Z,Chen X,Wang H,Qin L,Zhao W,Geng B

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Western Blot

Mon May 09 00:00:00 EDT 2022

Sulforaphane Protects Piglet Brains from Neonatal Hypoxic-Ischemic Injury.

Developmental neuroscience

Wang B,Kulikowicz E,Lee JK,Koehler RC,Yang ZJ

PA5-27882 was used in Western Blot to conclude that systemic administration of sulforaphane 15 min after HI can induce the translocation of Nrf2 to the nucleus, increase expression of an enzyme involved in glutathione synthesis, and salvage neurons in the highly vulnerable putamen and sensorimotor cortex in a large-animal model of HI.

Tue Oct 26 00:00:00 EDT 2021

The Role of Nrf2 Transcription Factor and Sp1-Nrf2 Protein Complex in Glutamine Transporter SN1 Regulation in Mouse Cortical Astrocytes Exposed to Ammonia.

International journal of molecular sciences

Dąbrowska K,Skowrońska K,Popek M,Albrecht J,Zielińska M

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Western Blot

Mon Oct 18 00:00:00 EDT 2021

Ficus erecta Thunb Leaves Alleviate Memory Loss Induced by Scopolamine in Mice via Regulation of Oxidative Stress and Cholinergic System.

Molecular neurobiology

Sohn E,Kim YJ,Kim JH,Jeong SJ

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Western Blot

Sun Aug 01 00:00:00 EDT 2021

P- Hydroxybenzyl Alcohol Alleviates Oxidative Stress in a Nonalcoholic Fatty Liver Disease Larval Zebrafish Model and a BRL-3A Hepatocyte Via the Nrf2 Pathway.

Frontiers in pharmacology

An J,Cheng L,Yang L,Song N,Zhang J,Ma K,Ma J

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Immunocytochemistry

Fri Apr 30 00:00:00 EDT 2021

Polarization of Low-Grade Inflammatory Monocytes Through TRAM-Mediated Up-Regulation of Keap1 by Super-Low Dose Endotoxin.

Frontiers in immunology

Rahtes A,Li L

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Immunoprecipitation

Mon Apr 19 00:00:00 EDT 2021

Polarization of Low-Grade Inflammatory Monocytes Through TRAM-Mediated Up-Regulation of Keap1 by Super-Low Dose Endotoxin.

Frontiers in immunology

Rahtes A,Li L

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Immunoprecipitation

Mon Apr 19 00:00:00 EDT 2021

Polarization of Low-Grade Inflammatory Monocytes Through TRAM-Mediated Up-Regulation of Keap1 by Super-Low Dose Endotoxin.

Frontiers in immunology

Rahtes A,Li L

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Immunoprecipitation

Mon Apr 19 00:00:00 EDT 2021

Biochemical study on the protective effect of curcumin on acetaminophen and gamma-irradiation induced hepatic toxicity in rats.

Environmental toxicology

Eassawy MMT,Salem AA,Ismail AFM

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Western Blot

Thu Dec 17 00:00:00 EST 2020

Chloroquine reverses chemoresistance via upregulation of p21^WAF1/CIP1 and autophagy inhibition in ovarian cancer.

Cell death & disease

Hwang JR,Kim WY,Cho YJ,Ryu JY,Choi JJ,Jeong SY,Kim MS,Kim JH,Paik ES,Lee YY,Han HD,Lee JW

PA5-27882 was used in Western Blotting to demonstrate that CQ increases cytotoxicity in combination with CDDP by inducing lethal DNA damage by induction of p21WAF1/CIP1 expression and autophagy inhibition in CDDP-resistant EOC.

Fri Dec 04 00:00:00 EST 2020

VEGFA and NFE2L2 Gene Expression and Regulation by MicroRNAs in Thyroid Papillary Cancer and Colloid Goiter.

Genes

Stuchi LP,Castanhole-Nunes MMU,Maniezzo-Stuchi N,Biselli-Chicote PM,Henrique T,Padovani Neto JA,de-Santi Neto D,Girol AP,Pavarino EC,Goloni-Bertollo EM

PA5-27882 was used in Immunohistochemistry to suggest that hsa-miR-17-5p may be a positive regulator of NFE2L2 expression in PTC.

Wed Aug 19 00:00:00 EDT 2020

Oxidative Stress Modulation by Cameroonian Spice Extracts in HepG2 Cells: Involvement of Nrf2 and Improvement of Glucose Uptake.

Metabolites

Nwakiban APA,Cicolari S,Piazza S,Gelmini F,Sangiovanni E,Martinelli G,Bossi L,Carpentier-Maguire E,Tchamgoue AD,Agbor G,Kuiaté JR,Beretta G,Dell'Agli M,Magni P

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Immunocytochemistry

Fri May 01 00:00:00 EDT 2020

NRF2 Activation Inhibits Both TGF-β1- and IL-13-Mediated Periostin Expression in Fibroblasts: Benefit of Cinnamaldehyde for Antifibrotic Treatment.

Oxidative medicine and cellular longevity

Mitamura Y,Murai M,Mitoma C,Furue M

PA5-27882 was used in Western Blotting to study the role of oxidative stress in the expression of POSTN induced by TGF-Β1 and IL-13 in dermal fibroblasts.

Wed Jan 02 00:00:00 EST 2019

Myokine irisin-induced protection against oxidative stress in vitro. Involvement of heme oxygenase-1 and antioxidazing enzymes superoxide dismutase-2 and glutathione peroxidase.

Journal of physiology and pharmacology : an official journal of the Polish Physiological Society

Mazur-Bialy AI,Kozlowska K,Pochec E,Bilski J,Brzozowski T

PA5-27882 was used in Western Blotting to determine the ability of irisin to interact with reactive oxygen species in RAW 264.7 murine macrophages.

Thu Feb 01 00:00:00 EST 2018

Acute maternal oxidant exposure causes susceptibility of the fetal brain to inflammation and oxidative stress.

Journal of neuroinflammation

Akhtar F,Rouse CA,Catano G,Montalvo M,Ullevig SL,Asmis R,Kharbanda K,Maffi SK

Published figure using Nrf2 polyclonal antibody (Product # PA5-27882) in Western Blot

Sat Sep 30 00:00:00 EDT 2017

Sodium fluoride affects zebrafish behaviour and alters mRNA expressions of biomarker genes in the brain: Role of Nrf2/Keap1.

Environmental toxicology and pharmacology

Mukhopadhyay D,Priya P,Chattopadhyay A

PA5-27882 was used in western blot to elucidate the molecular mechanism of fluoride-induced neurotoxicity using zebrafish

Tue Sep 01 00:00:00 EDT 2015

Acetyl-l-carnitine prevents homocysteine-induced suppression of Nrf2/Keap1 mediated antioxidation in human lens epithelial cells.

Molecular medicine reports

Yang SP,Yang XZ,Cao GP

PA5-27882 was used in western blot to prevent homocysteine-induced demethylation by administering acetyl-l-carnitine to human lens epithelial cells.

Wed Jul 01 00:00:00 EDT 2015

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: basic leucine zipper transcription factor