Overview
- Product nameAnti-NFkB p65 antibodySee all NFkB p65 primary antibodies ...
- DescriptionRabbit polyclonal to NFkB p65
- Specificityab31481 detects endogenous levels of NF-kB p65 protein around Serine 276.
- Tested applicationsICC/IF, WB, IHC-P, ELISA, IP more details
- Species reactivityReacts with: Mouse, Rat, Human, Pig
- Immunogen
Synthetic non-phosphopeptide derived from human NF-kB p65 around the phosphorylation site of Serine 276.
- Positive controlHeLa cells and breast carcinoma.
Properties
- FormLiquid
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 50% Glycerol, PBS (without Mg2+ and Ca2+), 150mM Sodium chloride, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Purification notesab31481 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
- Clonality Polyclonal
- IsotypeIgG
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Research Areas
Applications
Our Abpromise guarantee covers the use of ab31481 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| ICC/IF | ICC/IF: 1/200. See Abreview. |
| WB | WB: 1/500 - 1/1000. Detects a band of approximately 65 kDa. |
| IHC-P | IHC-P: Use at an assay dependent dilution. |
| ELISA | ELISA: 1/10000. |
| IP | IP: Use a concentration of 10 µg/ml. |
Target
- FunctionNF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1.
- Sequence similaritiesContains 1 RHD (Rel-like) domain.
- Domainthe 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
- Post-translational
modificationsUbiquitinated, leading to its proteasomal degradation. Degradation is required for termination of NF-kappa-B response.
Monomethylated at Lys-310 by SETD6. Monomethylation at Lys-310 is recognized by the ANK repeats of EHMT1 and promotes the formation of repressed chromatin at target genes, leading to down-regulation of NF-kappa-B transcription factor activity. Phosphorylation at Ser-311 disrupts the interaction with EHMT1 without preventing monomethylation at Lys-310 and relieves the repression of target genes.
Phosphorylation at Ser-311 disrupts the interaction with EHMT1 and promotes transcription factor activity (By similarity). Phosphorylation on Ser-536 stimulates acetylation on Lys-310 and interaction with CBP; the phosphorylated and acetylated forms show enhanced transcriptional activity.
Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3. Acetylation at Lys-122 enhances DNA binding and impairs association with NFKBIA. Acetylation at Lys-310 is required for full transcriptional activity in the absence of effects on DNA binding and NFKBIA association. Acetylation can also lower DNA-binding and results in nuclear export. Interaction with BRMS1 promotes deacetylation of 'Lys-310'. - Cellular localizationNucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalized with RELA in the nucleus upon TNF-alpha induction.
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Database links
- Entrez Gene: 5970 Human
- Entrez Gene: 19697 Mouse
- Entrez Gene: 309165 Rat
- Omim: 164014 Human
- SwissProt: Q04206 Human
- SwissProt: Q04207 Mouse
- Unigene: 502875 Human
- Unigene: 249966 Mouse
- Unigene: 19480 Rat
see all
Target information above from: UniProt accession
Q04206
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibodyMGC131774 antibodyNF kappa B p65delta3 antibody
- NFKB 3 antibodyNFKB3 antibodyNFKB3 antibodyNuclear Factor NF Kappa B p65 Subunit antibodyNuclear factor NF-kappa-B p65 subunit antibodyNuclear factor of kappa light polypeptide gene enhancer in B cells 3 antibodyNuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B Cells antibodyNuclear factor of kappa light polypeptide gene enhancer in B-cells 3 antibodyOTTHUMP00000233473 antibodyOTTHUMP00000233474 antibodyOTTHUMP00000233475 antibodyOTTHUMP00000233476 antibodyOTTHUMP00000233900 antibodyp65 antibodyp65 NF kappaB antibodyp65 NFkB antibodyRELA antibodyTF65_HUMAN antibodyTranscription Factor p65 antibodyv rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B cells 3 (p65)) antibodyV Rel Avian Reticuloendotheliosis Viral Oncogene Homolog A antibodyv rel reticuloendotheliosis viral oncogene homolog A (avian) antibodyV rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65 antibodyv-rel reticuloendotheliosis viral oncogene homolog A antibody
see all
Anti-NFkB p65 antibody images
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Immunohistochemistry (Paraffin-embedded sections) - NFkB p65 antibody (ab31481)ab31481 at 1/50, Immunohistochemical analysis of paraffin-embedded breast carcinoma. Left: Using ab31481, Right: The same antibody preincubated with synthesized peptide. -
Western blot - NFkB p65 antibody (ab31481)All lanes : Anti-NFkB p65 antibody (ab31481) at 1/500 dilution
Lane 1 : HeLa Cells (5 to 30ug of total protein).
Lane 2 : HeLa Cells (5 to 30ug of total protein). with The same antibody preincubated with synthesized peptide.
Observed band size : 65 kDa (why is the actual band size different from the predicted?) -
Immunocytochemistry/ Immunofluorescence - NFkB p65 antibody (ab31481)This image is courtesy of an anonymous Abreviewab31481 staining mouse BV-2 microglial cells by ICC/IF. Cells were fixed in 3:1 acetone:methanol and blocked in 2% Normal Goat Serum, 2% BSA, 0.3% Triton-X for 1 hour at RT. The primary antibody was diluted 1/200 and incubated with the sample for 2 hours. An Alexa Fluor® 488 conjugated goat anti-rabbit antibody, diluted 1/500, was used as the secondary.
Cells were plated on cover slips for 24 hour prior to staining and treated to show cytosolic followed by nuclear localization of p65.
References for Anti-NFkB p65 antibody (ab31481)
This product has been referenced in:
- Zampell JC et al. Temporal and spatial patterns of endogenous danger signal expression after wound healing and in response to lymphedema. Am J Physiol Cell Physiol 300:C1107-21 (2011). WB ; Mouse . Read more (PubMed: 21248077) »
- Jortay J et al. Local induction of adiponectin reduces lipopolysaccharide-triggered skeletal muscle damage. Endocrinology 151:4840-51 (2010). Read more (PubMed: 20702578) »
