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RabMAb

Anti-NFkB p100 抗体 [EPR4686] (ab109440)

製品の概要

  • 製品名Anti-NFkB p100 antibody [EPR4686]
    NFkB p100 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR4686] to NFkB p100
  • アプリケーション適用あり: ICC/IF, WB, IPmore details
    適用なし: Flow Cyt or IHC-P
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Synthetic peptide within Human NFkB p100 aa 700 to the C-terminus. The exact sequence is proprietary.
    Database link: Q00653

  • ポジティブ・コントロール
    • WB: Jurkat, HeLa, ECV-304, and MCF7 cell lysates ICC/IF: HAP1 cells and HAP1-NFkB p100/p52 knockout cells
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

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

    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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.
  • 追加情報Is unsuitable for Flow Cyt or IHC-P.
  • ターゲット情報

    • 関連性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.
    • 細胞内局在Cytoplasmic and Nuclear
    • 参照データベース
    • 別名
      • 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 antibody [EPR4686] 画像



    • 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 1/10000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 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 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?)

    Anti-NFkB p100 antibody [EPR4686] (ab109440) 使用論文

    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

    投稿 Oct 10 2013

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