Anti-NF-kB p65 [112A1021] antibody (ab13594)

Overview

  • Product nameAnti-NF-kB p65 [112A1021] antibodySee all NF-kB p65 primary antibodies ...
  • Description
    Mouse monoclonal [112A1021] to NF-kB p65
  • Tested applicationsWB, IHC-P, Flow Cyt more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Pig
  • Immunogen

    Synthetic peptide corresponding to Human NF-kB p65 aa 526-539.
    Sequence: GLLSGDEDFSSIAD

  • Positive control
    • Whole cell lysate from Daudi cells.
  • General notes

    This antibody binds to all forms of NF-KB p65, both activated (phosphorylated, dimers) and non-activated. NFkB in the nucleus is considered to be active.

Properties

Applications

Our Abpromise guarantee covers the use of ab13594 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 Use a concentration of 2 - 5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).
IHC-P Use a concentration of 5 µg/ml.
Flow Cyt Use 0.25-0.5µg for 106 cells.
  • Application notesIs unsuitable for ICC/IF.
  • 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 antibody
      • MGC131774 antibody
      • NF kappa B p65delta3 antibody
      • NFKB 3 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 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
      • v-rel reticuloendotheliosis viral oncogene homolog A antibody
      see all

    Anti-NF-kB p65 [112A1021] antibody images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human ovarian cystadenocarcinoma tissue labelling NF-kB p65 with ab13594 at 5µg/ml. Staining was enhanced by boiling tissue sections in 10mM sodium citrate buffer, pH6.0 for 10-20 minutes followed by cooling at room temperature for 20 minutes.

    • Flow Cytometric analysis of NF-kB p65 in HEK 293 cells labelled with ab13594 at 0.5µg (Red), or isotype control antibody (green).

    • All lanes : Anti-NF-kB p65 [112A1021] antibody (ab13594) at 2 µg/ml

      Lane 1 : Ramos cell lysate
      Lane 2 : Daudi cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : NIH 3T3 cell lysate

      Lysates/proteins at 30 µg per lane.


      Predicted band size : 65 kDa


    • Predicted band size : 65 kDa

      Western blot analysis of NF-kB p65 using ab13594 at 2µg/ml dilution against 10µg of Daudi cell lysate.

    References for Anti-NF-kB p65 [112A1021] antibody (ab13594)

    ab13594 has not yet been referenced specifically in any publications.

    Product Wall

    Application Western blot
    Sample Mouse Tissue lysate - whole (Lung)
    Loading amount 30 µg
    Specification Lung
    Gel Running Conditions Reduced Denaturing
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 21 2008

    Application Western blot
    Sample Human Cell lysate - whole cell (THP-1 cells)
    Loading amount 10 µg
    Specification THP-1 cells
    Gel Running Conditions Reduced Denaturing
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Sep 04 2007

    Thank you for contacting us for technical support with ab13594, I'm sorry the customer is disappointed with this product and I think I know the reasons for the problem and will detail below what I recommend. Our policy is to offer refunds or replaceme...

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