Anti-NFkB p100 / p52 antibody [EPR4686] (ab109440)

Overview

  • Product nameAnti-NFkB p100 / p52 antibody [EPR4686]
    See all NFkB p100 / p52 primary antibodies
  • Description
    Rabbit monoclonal [EPR4686] to NFkB p100 / p52
  • Tested applicationsSuitable for: ICC/IF, WB, IPmore details
    Unsuitable for: Flow Cyt or IHC-P
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment corresponding to residues in Human NFkB p100/ p52 (UniProt Q00653).

  • Positive control
    • Jurkat, HeLa, ECV-304, and MCF7 cell lysates ICC/IF: HAP1 cells and HAP1-NFkB p100/p52 knockout cells
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

Our Abpromise guarantee covers the use of ab109440 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 1/250.
WB 1/10000 - 1/50000. Detects a band of approximately 110 kDa (predicted molecular weight: 97 kDa).
IP 1/10 - 1/100.
  • Application notesIs unsuitable for Flow Cyt or IHC-P.
  • 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 [EPR4686] images



    • Predicted band size : 97 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: NFκB p100 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 - ab109440 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab109440 was shown to specifically react with NFκB p100 when NFκB p100 knockout samples were used. Wild-type and NFκB p100 knockout samples were subjected to SDS-PAGE. ab109440 and ab109440 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 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.

    • ab109440 staining NFkB p100/p52 in wild-type HAP1 cells (top panel) and NFkB p100/p52 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109440 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    • All lanes : Anti-NFkB p100 / p52 antibody [EPR4686] (ab109440) at 1/10000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : ECV-304 cell lysate
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 97 kDa
      Observed band size : 110 kDa (why is the actual band size different from the predicted?)

    References for Anti-NFkB p100 / p52 antibody [EPR4686] (ab109440)

    ab109440 has not yet been referenced specifically in any publications.

    Product Wall

    Application Western blot
    Loading amount 30 µg
    Gel Running Conditions Reduced Denaturing (Gel 8%)
    Sample Mouse Cell lysate - whole cell (WT & TRAF2 KO embryonic fibroblasts and macrophage)
    Specification WT & TRAF2 KO embryonic fibroblasts and macrophage
    Treatment TNFα, BV6, LPS and PAM-CSK4 for 24hrs
    Blocking step Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Ms. Anna McCormick

    Verified customer

    Submitted Oct 10 2013

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