Overview

  • Product nameAnti-NFkB p100 / p52 antibody [0.T.173]
    See all NFkB p100 / p52 primary antibodies
  • Description
    Mouse monoclonal [0.T.173] to NFkB p100 / p52
  • SpecificityThis antibody recognizes p52subunit of NFkB and its p100 precursor.
  • Tested applicationsSuitable for: WBmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Fusion protein (Human), corresponding to amino acids 1-444 of human NFkB and its p100 precursor.

  • Positive control
    • WB: Raji and HeLa nuclear extract cell lysate. No detection in mouse 3T3 cell and rat PC 12 cell lysates. IP: HeLa nuclear extract cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab32859 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
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 54 kDa.

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. 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.
  • Involvement in diseaseNote=A chromosomal aberration involving NFKB2 is found in a case of B-cell non Hodgkin lymphoma (B-NHL). Translocation t(10;14)(q24;q32) with IGHA1. The resulting oncogene is also called Lyt-10C alpha variant.
    Note=A chromosomal aberration involving NFKB2 is found in a cutaneous T-cell leukemia (C-TCL) cell line. This rearrangement produces the p80HT gene which encodes for a truncated 80 kDa protein (p80HT).
    Note=In B-cell leukemia (B-CLL) cell line, LB40 and EB308, can be found after heterogeneous chromosomal aberrations, such as internal deletions.
  • Sequence similaritiesContains 7 ANK repeats.
    Contains 1 death domain.
    Contains 1 RHD (Rel-like) domain.
  • DomainThe C-terminus of p100 might be involved in cytoplasmic retention, inhibition of DNA-binding by p52 homodimers, and/or transcription activation.
    The glycine-rich region (GRR) appears to be a critical element in the generation of p52.
  • Post-translational
    modifications
    While translation occurs, the particular unfolded structure after the GRR repeat promotes the generation of p52 making it an acceptable substrate for the proteasome. This process is known as cotranslational processing. The processed form is active and the unprocessed form acts as an inhibitor (I kappa B-like), being able to form cytosolic complexes with NF-kappa B, trapping it in the cytoplasm. Complete folding of the region downstream of the GRR repeat precludes processing.
    Subsequent to MAP3K14-dependent serine phosphorylation, p100 polyubiquitination occurs then triggering its proteasome-dependent processing.
    Constitutive processing is tightly suppressed by its C-terminal processing inhibitory domain, named PID, which contains the death domain.
  • Cellular localizationNucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor.
  • Information by UniProt
  • Database links
  • Alternative names
    • CVID10 antibody
    • DNA binding factor KBF2 antibody
    • DNA-binding factor KBF2 antibody
    • H2TF1 antibody
    • Lymphocyte translocation chromosome 10 antibody
    • Lymphocyte translocation chromosome 10 protein antibody
    • Lyt 10 antibody
    • Lyt10 antibody
    • NF kB2 antibody
    • NFKB2 antibody
    • NFKB2_HUMAN antibody
    • Nuclear factor NF kappa B p100 subunit antibody
    • Nuclear factor NF kappa B p52 subunit antibody
    • Nuclear factor NF-kappa-B p52 subunit antibody
    • Nuclear factor of kappa light chain gene enhancer in B cells 2 antibody
    • Nuclear factor of kappa light polypeptide gene enhancer in B cells 2 antibody
    • Nuclear factor of kappa light polypeptide gene enhancer in B-cells 2 antibody
    • Oncogene Lyt 10 antibody
    • Oncogene Lyt-10 antibody
    • p105 antibody
    • p49/p100 antibody
    see all

Anti-NFkB p100 / p52 antibody [0.T.173] images



  • Predicted band size : 54 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: NFκB p100/p52 knockout HAP1 cell lysate (20 µg)

    Lane 3: Jurkat cell lysate (20 µg)
    Lane 4: HeLa cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab32859 observed at 97 kDa. Red - loading control, ab181602, observed at 37 kDa.
    ab32859 was shown to recognize NFκB p100/p52 when NFκB p100/p52 knockout samples were used, along with additional cross-reactive bands. Wild-type and NFκB p100/p52 knockout samples were subjected to SDS-PAGE. ab32859 and ab181602 (loading control to NFκB p100/p52) were diluted 1 μg/mL and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Anti-NFkB p100 / p52 antibody [0.T.173] (ab32859) at 0.5 µg/ml + HeLa Cell nuclear extract

    Predicted band size : 54 kDa

References for Anti-NFkB p100 / p52 antibody [0.T.173] (ab32859)

This product has been referenced in:

See 1 Publication for this product

Product Wall

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Human Cell lysate - whole cell (B cells)
Specification B cells
Treatment CD40L for 48h
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Oct 29 2014

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