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RabMAbs (2229-1)

Anti-NFkB p65 antibody [EP2161Y] (ab76311)

Overview

  • Product nameAnti-NFkB p65 antibody [EP2161Y]See all NFkB p65 primary antibodies ...
  • Description
    Rabbit monoclonal [EP2161Y] to NFkB p65
  • Specificityab76311 detects both phosphorylated and non-phosphorylated versions of human NFkB p65.
  • Tested applicationsWB, ICC, Flow Cyt more details
  • Species reactivity
    Reacts with: Human

    Does not react with

    Mouse, Rat
  • Immunogen

    A synthetic phospho-peptide corresponding to residues surrounding serine 468 of human NFkB p65

  • Positive controlHeLa, Daudi and Jurkat cell lysates.
  • General notesProduced under U.S. Patent No. 5,675,063.

Applications

Our Abpromise guarantee covers the use of ab76311 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB WB: 1/100000 - 1/200000. Detects a band of approximately 70 kDa (predicted molecular weight: 65 kDa).
ICC ICC: 1/100 - 1/250.
Flow Cyt Flow Cyt: 1/30.
  • Application notesIs unsuitable for ,IHC-P or IP.
  • 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
      modifications
      Ubiquitinated, 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.
    • Target information above from: UniProt accession Q04206 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links
    • Alternative names
        Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibodyMGC131774 antibodyNF kappa B p65delta3 antibody
        NFKB 3 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 antibodyV rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65 antibody
      see all

    Anti-NFkB p65 antibody [EP2161Y] images

    • All lanes : Anti-NFkB p65 antibody [EP2161Y] (ab76311) at 1/20000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : Daudi cell lysate
      Lane 3 : Jurkat cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 65 kDa
      Observed band size : 70 kDa (why is the actual band size different from the predicted?)

    References for Anti-NFkB p65 antibody [EP2161Y] (ab76311)

    This product has been referenced in:
    • Jiao J  et al. The N terminus of adenovirus type 12 E1A inhibits major histocompatibility complex class I expression by preventing phosphorylation of NF-kappaB p65 Ser276 through direct binding. J Virol 84:7668-74 (2010). Read more (PubMed: 20504937) »

    See 1 Publication for this product

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    Thank you for contacting us. Flow cytometry protocols are recommended below. For most of the antibodies we are not able to provide specific epitope information but it seems most targets are intracellular. Hence not on the membrane and cells need fixat...

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