Key features and details
- Rabbit polyclonal to NF-kB p65 (phospho T254)
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Product nameAnti-NF-kB p65 (phospho T254) antibody
See all NF-kB p65 primary antibodies
DescriptionRabbit polyclonal to NF-kB p65 (phospho T254)
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide corresponding to Human NF-kB p65 conjugated to Keyhole Limpet Haemocyanin (KLH).
- 3T3 cell extract treated with TNF-alpha; HeLa cells
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.88% Sodium chloride
PBS is without Mg2+ and Ca2+
Concentration information loading...
PurityProtein A purified
Purification notesab131100 was purified by affinity chromatography using epitope specific phosphopeptide. Non phospho specific antibodies were removed by chromatography using non phosphopeptide.
Our Abpromise guarantee covers the use of ab131100 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/1000. Predicted molecular weight: 60 kDa.|
|IHC-P||1/50 - 1/100.|
|ICC/IF||1/100 - 1/200.|
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.
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.
- Information by UniProt
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All lanes : Anti-NF-kB p65 (phospho T254) antibody (ab131100) at 1/500 dilution
Lane 1 : 3T3 cell extract, untreated
Lane 2 : 3T3 cell extract treated with TNF-alpha
Predicted band size: 60 kDa
Actual band size : 65 kDa
Immunohistochemical analysis of NFkB p65(phospho T254) in paraffin-embedded human breast carcinoma tissue,using ab131100 at a dilution of 1/50.
Immunofluorescence analysis of NFkB p65 (phospho T254)in methanol fixed HeLa cells, using ab131100 at a dilution of 1/100.
ab131100 has been referenced in 2 publications.
- Wang Z et al. Polysaccharides from Enteromorpha Prolifera Ameliorate Acute Myocardial Infarction in Vitro and in Vivo via Up-Regulating HIF-1a. Int Heart J 60:964-973 (2019). PubMed: 31257333
- Li C et al. Swainsonine inhibits proliferation and collagen synthesis of NIH-3T3 cells by declining miR-21. Artif Cells Nanomed Biotechnol 47:2256-2264 (2019). PubMed: 31184213