Overview

  • Product nameAnti-NF-kB p65 (phospho S536) antibody
    See all NF-kB p65 primary antibodies
  • Description
    Rabbit polyclonal to NF-kB p65 (phospho S536)
  • Tested applicationsICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Guinea pig, Dog, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human NF-kB p65. around the phosphorylation site of Serine 536.

  • Positive control
    • Jurkat whole cell lysate treated with TNF alpha and Calyculin A. This antibody gave a positive result in IHC in the following FFPE tissue: human breast adenocarcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab86299 in the following tested applications.

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

Application Abreviews Notes
ICC/IF 1/50 - 1/250.
WB 1/2000 - 1/10000. Predicted molecular weight: 60 kDa.
IHC-P 1/100 - 1/500.

Epitope retrieval with Tris-EDTA pH9.0 is recommended

  • Application notesIs unsuitable for 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.
    • Information by UniProt
    • Database links
    • Alternative names
      • Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibody
      • MGC131774 antibody
      • NF kappa B p65delta3 antibody
      • NFKB3 antibody
      • Nuclear Factor NF Kappa B p65 Subunit antibody
      • Nuclear factor NF-kappa-B p65 subunit antibody
      • Nuclear factor of kappa light polypeptide gene enhancer in B cells 3 antibody
      • Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 antibody
      • OTTHUMP00000233473 antibody
      • OTTHUMP00000233474 antibody
      • OTTHUMP00000233475 antibody
      • OTTHUMP00000233476 antibody
      • OTTHUMP00000233900 antibody
      • p65 antibody
      • p65 NF kappaB antibody
      • p65 NFkB antibody
      • RELA antibody
      • TF65_HUMAN antibody
      • Transcription Factor p65 antibody
      • v rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B cells 3 (p65)) antibody
      • V rel avian reticuloendotheliosis viral oncogene homolog A antibody
      • v rel reticuloendotheliosis viral oncogene homolog A (avian) antibody
      • V rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65 antibody
      see all

    Anti-NF-kB p65 (phospho S536) antibody images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling NF-kB p65 (phospho S536) with ab86299 at 1/200 (1µg/ml) and 1/1000 (0.2µg/ml). Detection: DAB.
    • All lanes : Anti-NF-kB p65 (phospho S536) antibody (ab86299) at 0.04 µg/ml

      Lane 1 : Jurkat whole cell lysate treated with TNF alpha and Calyculin A
      Lane 2 : Jurkat whole cell lysate mock treated

      Lysates/proteins at 50 µg per lane.

      developed using the ECL technique

      Predicted band size : 60 kDa
      Observed band size : 70 kDa (why is the actual band size different from the predicted?)


      Exposure time : 30 seconds
    • IHC image of NF-kB p65 (phospho S536) staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab86299, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    References for Anti-NF-kB p65 (phospho S536) antibody (ab86299)

    This product has been referenced in:
    • López-Alonso I  et al. Impairment of autophagy decreases ventilator-induced lung injury by blockade of the NF-?B pathway. Am J Physiol Lung Cell Mol Physiol 304:L844-52 (2013). Mouse . Read more (PubMed: 23585228) »
    • Amado-Rodríguez L  et al. Anti-inflammatory effects of clarithromycin in ventilator-induced lung injury. Respir Res 14:52 (2013). WB ; Mouse . Read more (PubMed: 23663489) »

    See all 3 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 30 µg
    Gel Running Conditions Reduced Denaturing (10%)
    Sample Human Cell lysate - whole cell (LS174t)
    Specification LS174t
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 10 2014

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"