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RabMAb

Anti-NFkB p105 / p50抗体[E381] (ab32360)

概述

  • 产品名称
    Anti-NFkB p105 / p50抗体[E381]
    参阅全部 NFkB p105 / p50 一抗
  • 描述
    兔单克隆抗体[E381] to NFkB p105 / p50
  • 特异性
    This antibody will detect both forms: p50 and p105.
  • 经测试应用
    适用于: WB, IHC-Pmore details
    不适用于: Flow Cyt
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to NFkB p105/ p50 aa 300-400.

  • 阳性对照
    • WB: HeLa, MCF-7 and PC-12 cell lysates. IHC-P: human prostate carcinoma and bladder carcinoma tissues.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

     

    Alternative versions available:

    Anti-NFkB p105 / p50 antibody (Alexa Fluor® 647) [E381] (ab196519)

    Anti-NFkB p105 / p50 antibody (HRP) [E381] (ab195854)

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

性能

应用

Our Abpromise guarantee covers the use of ab32360 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000 - 1/10000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa).
IHC-P 1/250 - 1/500.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

  • 应用说明
    Is unsuitable for Flow Cyt.
  • 靶标

    • 功能
      NF-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 RelB-p50 complexes are transcriptional activators. The NF-kappa-B p50-p50 homodimer is a transcriptional repressor, but can act as a transcriptional activator when associated with BCL3. NFKB1 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p105 and generation of p50 by a cotranslational processing. The proteasome-mediated process ensures the production of both p50 and p105 and preserves their independent function, although processing of NFKB1/p105 also appears to occur post-translationally. p50 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. In a complex with MAP3K8, NFKB1/p105 represses MAP3K8-induced MAPK signaling; active MAP3K8 is released by proteasome-dependent degradation of NFKB1/p105.
    • 序列相似性
      Contains 7 ANK repeats.
      Contains 1 death domain.
      Contains 1 RHD (Rel-like) domain.
    • 结构域
      The C-terminus of p105 might be involved in cytoplasmic retention, inhibition of DNA-binding, and transcription activation.
      Glycine-rich region (GRR) appears to be a critical element in the generation of p50.
    • 翻译后修饰
      While translation occurs, the particular unfolded structure after the GRR repeat promotes the generation of p50 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.
      Phosphorylation at 'Ser-903' and 'Ser-907' primes p105 for proteolytic processing in response to TNF-alpha stimulation. Phosphorylation at 'Ser-927' and 'Ser-932' are required for BTRC/BTRCP-mediated proteolysis.
      Polyubiquitination seems to allow p105 processing.
      S-nitrosylation of Cys-61 affects DNA binding.
    • 细胞定位
      Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor.
    • Information by UniProt
    • 数据库链接
    • 别名
      • DKFZp686C01211 antibody
      • DNA binding factor KBF1 antibody
      • DNA binding factor KBF1 EBP1 antibody
      • DNA-binding factor KBF1 antibody
      • EBP 1 antibody
      • EBP-1 antibody
      • EBP1 antibody
      • KBF1 antibody
      • MGC54151 antibody
      • nf b antibody
      • NF kappa B antibody
      • NF kappaB antibody
      • NF kappabeta antibody
      • NF kB1 antibody
      • NFkappaB antibody
      • NFKB 1 antibody
      • NFKB p105 antibody
      • NFKB p50 antibody
      • Nfkb1 antibody
      • NFKB1_HUMAN antibody
      • Nuclear factor kappa B DNA binding subunit antibody
      • Nuclear factor kappa-B, subunit 1 antibody
      • Nuclear factor NF kappa B p105 subunit antibody
      • Nuclear factor NF kappa B p50 subunit antibody
      • Nuclear factor NF-kappa-B p50 subunit antibody
      • Nuclear factor of kappa light chain gene enhancer in B cells 1 antibody
      • Nuclear factor of kappa light polypeptide gene enhancer in B cells 1 antibody
      • Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 antibody
      • p105 antibody
      • p50 antibody
      • p84/NF-kappa-B1 p98 antibody
      • Transcription factor NFKB1 antibody
      see all

    图片



    • Predicted band size : 50 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: NFkB p105 / p50 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Jurkat cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab32360 observed at 120, 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab32360 was shown to specifically react with NFkB p105 / p50 when NFkB p105 / p50 knockout samples were used. Wild-type and NFkB p105 / p50 knockout samples were subjected to SDS-PAGE. ab32360 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and 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 hour at room temperature before imaging.

    • Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified) + HeLa cell lysate at 20 µg

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 50 kDa
      Observed band size : 50,105 kDa (why is the actual band size different from the predicted?)

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling NFkB p105 / p50 with purified ab32360 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/50000 dilution (purified) + MCF-7 cell lysate at 10 µg

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 50 kDa
      Observed band size : 105 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 50 kDa (possible non-specific binding).

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified)

      Lane 1 : PC-12 cell lysate
      Lane 2 : NIH/3T3 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 50 kDa
      Observed band size : 105 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 50 kDa (possible non-specific binding).

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/5000 dilution (unpurified) + HeLa cell lysate

      Predicted band size : 50 kDa
      Observed band size : 50,105 kDa (why is the actual band size different from the predicted?)
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling NFkB p105 / p50 with unpurified ab32360 at a dilution of 1/250.

    实验方案

    文献

    This product has been referenced in:
    • Aroune D  et al. Changes in the NF?B and E-cadherin expression are associated to diabetic nephropathy inPsammomys obesus. Saudi J Biol Sci 24:843-850 (2017). IHC ; Sand rat . Read more (PubMed: 28490956) »
    • Fei L  et al. Glycyrrhizin Ameliorate Ischemia Reperfusion Lung Injury through Downregulate TLR2 Signaling Cascade in Alveolar Macrophages. Front Pharmacol 8:389 (2017). WB ; Mouse . Read more (PubMed: 28670282) »

    See all 37 Publications for this product

    客户评价及客户问答

    Thank you for contacting us. Flow cytometry protocols are recommended below. For most of the antibodies we are not able to provide specific epitope information but it seems most targets are intracellular. Hence not on the membrane and cells need fi...

    Read More

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

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