Invitrogen

Phospho-NFkB p65 (Ser529) Monoclonal Antibody (B33B4WP), eFluor™ 450, eBioscience™

1 Published Figure
1 Reference
View all (154) NFkB p65 antibodies
Datasheet
Protocols
Questions & Answers
Datasheet
Protocols
Questions & Answers

Product Details

48-9863-42

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.125 µg)/test
View 1 publication 1 publication
Product Specifications

Species Reactivity

Human

Published species

Not Applicable

Host/Isotype

Mouse / IgG2a, kappa

Recommended Isotype Control

Mouse IgG2a kappa Isotype Control (eBM2a), eFluor™ 450, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

B33B4WP

Conjugate

eFluor™ 450 eFluor™ 450 eFluor™ 450

Excitation/Emission Max

405/445 nm View spectra spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2815188

Product Specific Information

Description: The B33B4WP monoclonal antibody recognizes human NF kappa B (NFkB) p65 subunit when serine 529 is phosphorylated. NFkB, also known as nuclear factor kappa-light chain enhancer of activated B cells, is a ubiquitous transcription factor that regulates the transcription of many genes involved in cell proliferation, apoptosis, development, immunity, and cancer. Functional NFkB is a homo- or heterodimer composed of 5 members of the NFkB family: p65 (RelA), c-Rel, RelB, p50 (NFkB1, p105 precursor protein), and p52 (NFkB2, p100 precursor protein). The activity of the complex is negatively regulated by binding to IkB inhibitors that sequester NFkB into the cytoplasm, inhibiting its transcriptional activity. NFkB-activating agents like tumor necrosis factor (TNF) alpha, interleukin-1 beta, lipopolysaccharide, camptothecin, and phorbol ester (PMA) induce the phosphorylation and degradation of IkB, leading to the translocation of NFkB to the nucleus where it binds to kB motifs and regulates gene expression. The activity of p65-containing NFkB complexes is positively regulated by phosphorylation of the p65 subunit at serine 529 and/or serine 536.

The B33B4WP monoclonal antibody recognizes a single band at 65 kDa in reduced SDS lysates from TNF alpha-stimulated HeLa cells.

Applications Reported: This B33B4WP antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This B33B4WP antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol A: Two-step protocol for intracellular (cytoplasmic) proteins" located at www.thermofisher.com/flowprotocols . This may be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

eFluor 450 is an alternative to Pacific Blue. eFluor 450 emits at 446 nm and is excited with the violet laser line (405 nm). Please make sure that your instrument is capable of detecting this fluorochrome.

Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser.

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.

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

Bioinformatics

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

View more View less

Gene Aliases: NFKB3; p65; RELA

View more View less

UniProt ID: (Human) Q04206

View more View less

Entrez Gene ID: (Human) 5970

View more View less

Function(s)
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 Rel homology transcription factor
Process(es)
negative regulation of transcription from RNA polymerase II promoter liver development hair follicle development stimulatory C-type lectin receptor signaling pathway acetaldehyde metabolic process transcription from RNA polymerase II promoter inflammatory response cellular defense response aging positive regulation of cell proliferation organ morphogenesis response to organic substance response to UV-B positive regulation of Schwann cell differentiation cytokine-mediated signaling pathway membrane protein intracellular domain proteolysis positive regulation of chondrocyte differentiation positive regulation of type I interferon production response to muramyl dipeptide response to progesterone response to insulin response to cobalamin cellular response to hepatocyte growth factor stimulus response to muscle stretch Fc-epsilon receptor signaling pathway negative regulation of protein catabolic process response to drug negative regulation of apoptotic process positive regulation of I-kappaB kinase/NF-kappaB signaling response to amino acid response to morphine positive regulation of interleukin-12 biosynthetic process negative regulation of transcription, DNA-templated positive regulation of transcription, DNA-templated positive regulation of transcription from RNA polymerase II promoter negative regulation of insulin receptor signaling pathway regulation of inflammatory response T cell receptor signaling pathway positive regulation of NF-kappaB transcription factor activity response to cAMP defense response to virus cellular response to hydrogen peroxide nucleotide-binding oligomerization domain containing 2 signaling pathway response to interleukin-1 cellular response to lipopolysaccharide cellular response to nicotine cellular response to interleukin-1 cellular response to interleukin-6 cellular response to tumor necrosis factor cellular response to peptide hormone stimulus negative regulation of NIK/NF-kappaB signaling positive regulation of miRNA metabolic process negative regulation of extrinsic apoptotic signaling pathway
guarantee_icon

Performance Guarantee

If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*

Learn more
help_icon

We're here to help

Get expert recommendations for common problems or connect directly with an on staff expert for technical assistance related to applications, equipment and general product use.

Contact tech support