Invitrogen

NFkB p65 Polyclonal Antibody, eBioscience™

Advanced Verification

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

10 Published Figures

18 References

View all (144) NFkB p65 antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

NFkB p65 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Transcription factor p65 was performed using HeLa cells (serum-starved) and HeLa cells serum-starved for 16 hours, followed by 20 ng/mL human TNF alpha treatment for 40 minutes. 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 NFkB p65 Polyclonal Antibody, eBioscience™ (Product # 14-6731-81) 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 localization of p65 protein in HeLa treated cells. Panel e represents the merged image of HeLa untreated cells that shows cytoplasmic localization of p65 protein. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

NFkB p65 Antibody in Western Blot (WB) — Western Blot was performed using Anti-NFkB p65 Polyclonal Antibody, eBioscience™ (Product # 14-6731-81) and a 60 kDa band corresponding to Transcription factor p65 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), SK-BR-3 (Lane 2), NIH/3T3 (Lane 3), PC-12 (Lane 4) 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 (1:2000 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

NFkB p65 Antibody in ChIP Assay (ChIP) — Chromatin Immunoprecipitation (ChIP) assay of endogenous NFkB p65 was performed using NFkB p65 Polyclonal Antibody, eBioscience™ (Product # 14-6731-81, 5 µg) on sheared chromatin from HeLa cells treated with TNF-alpha (50 ng/mL for 1hr) using the MAGnify ChIP System kit (Product # 49-2024). Chromatin from untreated HeLa cells were prepared the same way. Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to IL8, IKKB, MYOD and SAT alpha (inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

NFkB p65 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Published figure using NFkB p65 polyclonal antibody (Product # 14-6731-81) in Immunohistochemistry (Paraffin)

NFkB p65 Antibody in Immunohistochemistry (IHC) — Fig. 2 TLR7 pathway is activated on stimulation with its specific ligand imiquimod, R837. Chief downstream regulators, especially NF-kB is activated and transclocated to the nucleus on TLR7 activation. a Host immune response on TLR7 activation, using TLR7 ligand. HepG2.2.15 cells were treated with TLR7 agonist, R837 (8mug/ml) for 3 days. mRNA expression of different downstream regulators (NF- KB, JNK, p38 and PI3K) involved in the TLR7 pathway. b Confocal imaging showing nuclear uptake of Nuclear Factor-KB p65 subunit (NF- KB) from cytoplasm in cells where HepG2.2.15 cells are taken as control and HepG2.2.15 cells stimulated with TLR7 ligand, R837 (8mug/ml) for 72 h. c Nuclear Factor-KB p65 subunit (NF-KB) protein expression of HepG2 and HepG2.2.15 cells. d Nuclear Factor-KB p65 subunit (NF- KB) protein expression of HepG2.2.15 cells taken as control and treated with R837 (8mug/ml) for 72 h

NFkB p65 Antibody in Western Blot, Gel Shift (WB, GS) — Fig. 1 No transcriptional NF-kappaB activity after PrP Sc infection. A. Electromobility shift assay revealed NF-kappaB binding activity in PrP Sc -infected neuroblastoma cells (Bos2). Nuclear extracts of Bos2 cells infected with RML6, treated with CD1 or stimulated with 10 ng ml -1 TNF-alpha were incubated with 32 P-labelled double-stranded oligonucleotide specific for NF-kappaB. The TNF-alpha sample was incubated with a 50-fold excess of cold NF-kappaB oligonucleotide as specific competitor or with a mutated oligonucleotide as control. B. kappaB precipitation experiment and detection using antibodies directed against p65, p50, p52 and Bcl-3. kappaB binding proteins were precipitated from nuclear extracts using an agarose-conjugated consensus kappaB oligonucleotide. The precipitates from either untreated controls, TNF-treated or RML6-infected Bos2 cells were divided and analysed in Western blots for the presence of the various NF-kappaB subunits. The immunoblot was quantified by scanning densitometry. The total signal intensities (p65 + p50, +p52 + Bcl-3) for the treated cells relative to the control cells and the portion of each subunits are given in percentage. C. Luciferase reporter gene assay showed no transcriptional NF-kappaB activity after PrP Sc infection (0.2% or 0.02%), the value of relative light unit (RLU) per mug protein being equivalent for RML6- and CD1-treated cells. Mean values and SEM of three independent transfections are shown.

NFkB p65 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with Transcription factor p65 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-NFkB p65 Polyclonal Antibody, eBioscience™ (Product # 14-6731-81). {KD}

Product Details

14-6731-81

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000-1:2,000	View 8 publications 8 publications
Immunohistochemistry (IHC)	1:1,000	View 3 publications 3 publications
Immunocytochemistry (ICC/IF)	1:100	View 3 publications 3 publications
Immunoprecipitation (IP)	Assay-Dependent	-
ChIP assay (ChIP)	2.5 µg/10^6 cells	View 1 publication 1 publication
Immunomicroscopy (IM)	-	View 1 publication 1 publication
Immunohistochemistry (Paraffin) (IHC (P))	-	View 1 publication 1 publication
Gel Shift (GS)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Conjugate	Unconjugated
Form	Liquid
Concentration	0.2 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS
Contains	0.1% sodium azide
Storage conditions	4° C
RRID	AB_468255

Product Specific Information

Description: The polyclonal antibody reacts with mouse, rat, and human NFkB p65; the antibody was raised against a peptide mapping to the carboxy terminus of human NFkB p65. Members of the rel/NFkB family of transcription factors are involved in the regulation of cellular responses, such as growth, development, and the inflammatory response. They share a structural motif known as the rel homology region (RHR), the C-terminal one third of which mediates protein dimerization (2, 6, 8). Complexes of p50 (NF-kB1) or p52 (NF-kB2) are generated through the processing of p105 and p100 precursors, respectively. These are usually associated with members of the Rel family (p65, c-Rel, Rel B). The homo- and heterodimer formed through combinations of NFkB/Rel proteins bind distinct kB sites to regulate the transcription of different genes (7, 9). In resting cells, NFkB is retained in the cytoplasm bound to inhibitory proteins of the IkB family. Degradation of IkB proteins occurs with cell activation, via of variety of signals, including inflammatory cytokines and bacterial lipopolysaccharides (LPS) as well as oxidative and fluid mechanical stress. This results in nuclear translocation of NFkB and the transcriptional gene activation of proinflammatory genes (1, 9). It has been suggested that NFkB plays a role in the development of numerous pathological states. Activation of NFkB induces gene programs leading to transcription of factors that promote inflammation, such as leukocyte adhesion molecules, cytokines, and chemokines. It is also thought that there are some substances with possible anti-inflammatory effects that are also NFkB regulated. There is some evidence indicating NFkB as a key factor in the pathophysiology of cardiac ischemia-reperfusion injury as well as the development of insulin dependent Diabetes Mellitus (4, 3).

Applications Reported: Purified anti-mo/hu/rat NFkB p65 poly has been reported for use in immunoprecipitation, immunoblotting (WB), and immunohistochemical staining.

Applications Tested: Purified anti-mo/hu/rat NFkB p65 poly has been tested by immunoblotting (WB) and immunohistochemistry. (1:1000 starting dilution). It is recommended that this antibody be titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Nuclear factor kB p65 (NF-kB p65) is encoded by the RELA gene and is present on chromosome 11 in humans. NF-kB P65 is also known as RelA (v-rel avian reticuloendotheliosis viral oncogene homolog A) and belongs to the Rel family of proteins. It is one of the two subunits of NF-kB (Nuclear factor kappa-light-chain-enhancer of activated B cells) that heterodimerizes with the other subunits p50 or p52. However, binding of TNF alpha to its cognate receptor phosphorylates IKK which in turn phosphorylates I kB allowing proteasomal degradation of I kB. It specifically plays a key role in transcription of immunoglobulin k (kappa) gene in mature B-lymphoid cells.

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

Product References

Malaria parasite heme biosynthesis promotes and griseofulvin protects against cerebral malaria in mice.

Nature communications

Chandana M,Anand A,Ghosh S,Das R,Beura S,Jena S,Suryawanshi AR,Padmanaban G,Nagaraj VA

14-6731-81 was used in Western Blotting to show heme synthesized in asexual stages promotes cerebral pathogenesis by enhancing hemozoin formation.

Tue Jul 12 00:00:00 EDT 2022

Inhibitory effects of quail egg on mast cells degranulation by suppressing PAR2-mediated MAPK and NF-kB activation.

Food & nutrition research

Lianto P,Ogutu FO,Zhang Y,He F,Che H

14-6731 was used in Western Blotting to investigate the suppressive role of quail egg in mast cell degranulation and cytokine production of the effect phase response.

Fri Mar 18 00:00:00 EDT 2022

Butorphanol reduces the neuronal inflammatory response and apoptosis via inhibition of p38/JNK/ATF2/p53 signaling.

Experimental and therapeutic medicine

Huang Y,Li S,Chen H,Feng L,Yuan W,Han T

Published figure using NFkB p65 polyclonal antibody (Product # 14-6731-81) in Western Blot

Tue Mar 01 00:00:00 EST 2022

Modulation of allergic inflammation in the lung by a peptide derived from Mycobacteria tuberculosis chaperonin 60.1.

Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

Riffo-Vasquez Y,Kanabar V,Keir SD,E-Lacerda RR,Man F,Jackson DJ,Corrigall V,Coates ARM,Page CP

14-6731 was used in Immunohistochemistry to investigate the effect of a shorter peptide sequence derived from Cpn 60.1, named IRL201104, on allergic lung inflammation induced by ovalbumin (OVA) in mice and by house dust mite (HDM) in guinea pigs, as well as investigating the action of IRL201104 on human cells in vitro.

Wed Apr 01 00:00:00 EDT 2020

Palmitic acid induces guanylin gene expression through the Toll-like receptor 4/nuclear factor-κB pathway in rat macrophages.

American journal of physiology. Cell physiology

Akieda-Asai S,Ma H,Date Y

14-6731 was used in Chromatin immunoprecipitation to indicate that saturated fatty acids, such as palmitic acid, activate guanylin gene expression via the nuclear factor-κB pathway in macrophages.

Sun Dec 01 00:00:00 EST 2019

Anti-inhibitory potential of an ethanolic extract of Distromium decumbens on pro-inflammatory cytokine production in Pseudomonas aeruginosa lipopolysaccharide-stimulated nasal polyp-derived fibroblasts.

International journal of molecular medicine

Lee DS,Lee CM,Park SK,Yim MJ,Lee JM,Choi G,Yoo JS,Jung WK,Park S,Seo SK,Park WS,Choi IW

Fri Dec 01 00:00:00 EST 2017

Free immunoglobulin light chain (FLC) promotes murine colitis and colitis-associated colon carcinogenesis by activating the inflammasome.

Scientific reports

Ma J,Jiang D,Gong X,Shao W,Zhu Z,Xu W,Qiu X

14-6731 was used in Western Blotting to investigate whether free Ig light chain is involved in colitis and colitis-associated colon carcinogenesis.

Wed Jul 12 00:00:00 EDT 2017

Anti-hepatitis B virus (HBV) response of imiquimod based toll like receptor 7 ligand in hbv-positive human hepatocelluar carcinoma cell line.

BMC infectious diseases

Das D,Sengupta I,Sarkar N,Pal A,Saha D,Bandopadhyay M,Das C,Narayan J,Singh SP,Chakrabarti S,Chakravarty R

Published figure using NFkB p65 polyclonal antibody (Product # 14-6731-81) in Immunofluorescence

Sat Jan 14 00:00:00 EST 2017

Down-regulation of IKKβ expression in glioma-infiltrating microglia/macrophages is associated with defective inflammatory/immune gene responses in glioblastoma.

Oncotarget

Mieczkowski J,Kocyk M,Nauman P,Gabrusiewicz K,Sielska M,Przanowski P,Maleszewska M,Rajan WD,Pszczolkowska D,Tykocki T,Grajkowska W,Kotulska K,Roszkowski M,Kostkiewicz B,Kaminska B

14-6731 was used in Immunocytochemistry to show that down-regulation of IKBKB and NFκB signaling in microglia/macrophages infiltrating glioblastoma correlates with defective expression of immune/inflammatory genes and M2 polarization.

Tue Oct 20 00:00:00 EDT 2015

YCG063 inhibits Pseudomonas aeruginosa LPS-induced inflammation in human retinal pigment epithelial cells through the TLR2-mediated AKT/NF-κB pathway and ROS-independent pathways.

International journal of molecular medicine

Paeng SH,Park WS,Jung WK,Lee DS,Kim GY,Choi YH,Seo SK,Jang WH,Choi JS,Lee YM,Park S,Choi IW

14-6731-81 was used in Western Blot to suggest that this anti-inflammatory activity occurred through ROS-independent signaling pathways.

Tue Sep 01 00:00:00 EDT 2015

3,3'-Diindolylmethane inhibits VEGF expression through the HIF-1α and NF-κB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions.

International journal of molecular medicine

Park H,Lee DS,Yim MJ,Choi YH,Park S,Seo SK,Choi JS,Jang WH,Yea SS,Park WS,Lee CM,Jung WK,Choi IW

14-6731 was used in Electrophoretic mobility shift assay to demonstrate that DIM inhibits the CoCl2-induced production of VEGF by suppressing mitochondrial ROS production.

Wed Jul 01 00:00:00 EDT 2015

Transmembrane peptides used to investigate the homo-oligomeric interface and binding hotspot of latent membrane protein 1.

Biopolymers

Sammond DW,Joce C,Takeshita R,McQuate SE,Ghosh N,Martin JM,Yin H

Tue Nov 01 00:00:00 EDT 2011

Knockout of the tumor suppressor gene Gprc5a in mice leads to NF-kappaB activation in airway epithelium and promotes lung inflammation and tumorigenesis.

Cancer prevention research (Philadelphia, Pa.)

Deng J,Fujimoto J,Ye XF,Men TY,Van Pelt CS,Chen YL,Lin XF,Kadara H,Tao Q,Lotan D,Lotan R

Thu Apr 01 00:00:00 EDT 2010

Knockout of the tumor suppressor gene Gprc5a in mice leads to NF-kappaB activation in airway epithelium and promotes lung inflammation and tumorigenesis.

Cancer prevention research (Philadelphia, Pa.)

Deng J,Fujimoto J,Ye XF,Men TY,Van Pelt CS,Chen YL,Lin XF,Kadara H,Tao Q,Lotan D,Lotan R

Thu Apr 01 00:00:00 EDT 2010

Knockout of the tumor suppressor gene Gprc5a in mice leads to NF-kappaB activation in airway epithelium and promotes lung inflammation and tumorigenesis.

Cancer prevention research (Philadelphia, Pa.)

Deng J,Fujimoto J,Ye XF,Men TY,Van Pelt CS,Chen YL,Lin XF,Kadara H,Tao Q,Lotan D,Lotan R

Thu Apr 01 00:00:00 EDT 2010

NF-kappaB activation through the alternative pathway correlates with chemoresistance and poor survival in extranodal NK/T-cell lymphoma, nasal type.

Japanese journal of clinical oncology

Liu X,Wang B,Ma X,Guo Y

14-6731 was used in Immunohistochemistry to investigate how NF-kappaB activation through the alternative pathway is frequently observed in NK/T-cell lymphoma and associated with chemoresistance and poor survival.

Wed Jul 01 00:00:00 EDT 2009

Alteration of NF-kappaB activity leads to mitochondrial apoptosis after infection with pathological prion protein.

Cellular microbiology

Bourteele S,Oesterle K,Weinzierl AO,Paxian S,Riemann M,Schmid RM,Planz O

Sat Sep 01 00:00:00 EDT 2007

Alteration of NF-kappaB activity leads to mitochondrial apoptosis after infection with pathological prion protein.

Cellular microbiology

Bourteele S,Oesterle K,Weinzierl AO,Paxian S,Riemann M,Schmid RM,Planz O

Sat Sep 01 00:00:00 EDT 2007

Bioinformatics

Protein Aliases: avian reticuloendotheliosis viral (v-rel) oncogene homolog A; MGC131774; NF KB; nf-kappa-b p65; NF-kappa-B p65delta3; NF-kappa-B transcription factor p65; NFkappaB p65; nfkb rela; NFkB-p50; NFKB1; nuclear factor; nuclear factor kappa b; nuclear factor kappa B subunit p65; Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3; p65 NF kappaB; p65 NF-kappa B; p65 NFkB; REL A; Transcription factor p65; v-rel avian reticuloendotheliosis viral oncogene homolog A; v-rel reticuloendotheliosis viral oncogene homolog A

Gene Aliases: NFkB; NFKB3; p65; RELA

UniProt ID: (Human) Q04206, (Mouse) Q04207

Entrez Gene ID: (Human) 5970, (Mouse) 19697, (Rat) 309165

Molecular Function: Rel homology transcription factor

Function(s)

Process(es)

RNA polymerase II regulatory region sequence-specific DNA binding RNA polymerase II core promoter proximal region sequence-specific DNA binding RNA polymerase II distal enhancer sequence-specific DNA binding transcriptional activator activity, RNA polymerase II core promoter proximal region sequence-specific binding transcriptional repressor activity, RNA polymerase II core promoter proximal region sequence-specific binding transcriptional activator activity, RNA polymerase II distal enhancer sequence-specific binding DNA binding chromatin binding transcription factor activity, sequence-specific DNA binding transcription factor activity, RNA polymerase II distal enhancer sequence-specific binding protein binding transcription factor binding protein kinase binding chromatin DNA binding ubiquitin protein ligase binding protein complex binding activating transcription factor binding phosphate ion binding identical protein binding protein homodimerization activity actinin binding histone deacetylase binding transcription regulatory region DNA binding protein heterodimerization activity protein N-terminus binding NF-kappaB binding repressing transcription factor binding ankyrin repeat binding enzyme binding sequence-specific DNA binding RNA polymerase II transcription factor activity, sequence-specific DNA binding

NFkB p65 Polyclonal Antibody, eBioscience™

Product Details

14-6731-81

Western Blot (WB)

Immunohistochemistry (IHC)

Immunocytochemistry (ICC/IF)

Immunoprecipitation (IP)

ChIP assay (ChIP)

Immunomicroscopy (IM)

Immunohistochemistry (Paraffin) (IHC (P))

Gel Shift (GS)

Species Reactivity

Published species

Host/Isotype

Class

Type

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

Performance Guarantee

We're here to help

Your items have has been added!