Anti-NFkB p100 (phospho S865) antibody (ab31474)

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Overview

  • Product name
    Anti-NFkB p100 (phospho S865) antibody
    See all NFkB p100 primary antibodies
  • Description
    Rabbit polyclonal to NFkB p100 (phospho S865)
  • Specificity
    This antibody detects endogenous levels of NFkB p100 protein only when phosphorylated at Serine 865.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, ELISA, IPmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human NFkB p100 (phospho S865). The exact sequence is proprietary.

  • Positive control
    • WB: extracts of ovary cancer cells. IHC-P:breast carcinoma tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab31474 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 Use a concentration of 1 µg/ml.
WB 1/500 - 1/1000. Predicted molecular weight: 54 kDa.
IHC-P 1/50 - 1/100.
ELISA 1/20000.
IP Use at an assay dependent concentration.

Target

  • Relevance
    NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes 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. 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. In a non-canonical activation pathway, the MAP3K14-activated CHUK/IKKA homodimer phosphorylates NFKB2/p100 associated with RelB, inducing its proteolytic processing to NFKB2/p52 and the formation of NF-kappa-B RelB-p52 complexes. The NF-kappa-B heterodimeric RelB-p52 complex is a transcriptional activator. The NF-kappa-B p52-p52 homodimer is a transcriptional repressor. NFKB2 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p100 and generation of p52 by a cotranslational processing. The proteasome-mediated process ensures the production of both p52 and p100 and preserves their independent function. p52 binds to the kappa-B consensus sequence 5'-GGRNNYYCC-3', located in the enhancer region of genes involved in immune response and acute phase reactions. p52 and p100 are respectively the minor and major form; the processing of p100 being relatively poor. Isoform p49 is a subunit of the NF-kappa-B protein complex, which stimulates the HIV enhancer in synergy with p65. In concert with RELB, regulates the circadian clock by repressing the transcriptional activator activity of the CLOCK-ARNTL/BMAL1 heterodimer.
  • Cellular localization
    Cytoplasmic and Nuclear
  • Database links
    see all
  • Alternative names
    • CVID10 antibody
    • DNA binding factor KBF2 antibody
    • H2TF1 antibody
    • Lymphocyte translocation chromosome 10 protein antibody
    • LYT 10 antibody
    • NF kB2 antibody
    • NFKB p52/p100 subunit antibody
    • Nuclear factor Kappa B subunit 2 antibody
    • Nuclear factor of kappa light polypeptide gene enhancer in B cells 2 (p49/p100) antibody
    • Nuclear factor of kappa light polypeptide gene enhancer in B cells 2 antibody
    • Oncogene Lyt 10 antibody
    • p100 antibody
    • Transcription factor NFKB2 antibody
    see all

Anti-NFkB p100 (phospho S865) antibody images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p100 (phospho S865) antibody (ab31474)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p100 (phospho S865) antibody (ab31474)

    Ab31474, at a dilution of 1/50, staining NFkB p100 in paraffin embedded human breast carcinoma tissue by Immunohistochemistry.
    Left image: treated with ab31474.
    Right image: treated with the same antibody preincubated with sythesized peptide.

  • Western blot - Anti-NFkB p100 (phospho S865) antibody (ab31474)
    Western blot - Anti-NFkB p100 (phospho S865) antibody (ab31474)
    All lanes : Anti-NFkB p100 (phospho S865) antibody (ab31474) at 1/500 dilution

    Lane 1 : Extracts of ovary cancer cells (5-30ug).
    Lane 2 : Extracts of ovary cancer cells (5-30ug). Antibody pre-incubated with synthesized peptide.

    Secondary
    goat-anti-rabbit IgG-AP-conjugate.

    Predicted band size : 54 kDa
  • Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100 (phospho S865) antibody (ab31474)
    Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100 (phospho S865) antibody (ab31474)
    ICC/IF image of ab31474 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab31474, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-NFkB p100 (phospho S865) antibody (ab31474)

ab31474 has not yet been referenced specifically in any publications.

Publishing research using ab31474? Please let us know so that we can cite the reference in this datasheet.

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1 Abreview or Q&A

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Votes: Highest First

What staining pattern is seen with this antibody in normal tissue (from the image on the datasheet, it appears tumor tissue is nuclear)? Does this antibody stain mitochondrial DNA?

Thank you for your enquiry. The originator had their QC group do IHC and dot blot (DNA) for your questions. The data showed that 1) main staining was in nuclear for both tumor and normal tissues used, and 2) This antibody did not react with mitochon...

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