製品の概要

  • 製品名Anti-IKB alpha antibody [EP697]
    IKB alpha 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP697] to IKB alpha
  • アプリケーション適用あり: WB, Flow Cyt, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human IKB alpha (C terminal).

  • ポジティブ・コントロール
    • HeLa, PC-12, RAW264.7 and Jurkat cell lysates; human kidney and colon tissue.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody. 

    Alternative versions available:
    Anti-IKB alpha antibody (Alexa Fluor® 647) [EP697] (ab207356)

    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.

    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.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab76429 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
WB 1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
Flow Cyt 1/250.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/1000.

IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

For unpurified use at 1/100 - 1/250.

ICC/IF 1/250.

For unpurified use at 1/50 - 1/100.

ターゲット情報

  • 機能Inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription.
  • 関連疾患Ectodermal dysplasia, anhidrotic, with T-cell immunodeficiency autosomal dominant
  • 配列類似性Belongs to the NF-kappa-B inhibitor family.
    Contains 5 ANK repeats.
  • 翻訳後修飾Phosphorylated; disables inhibition of NF-kappa-B DNA-binding activity. Phosphorylation at positions 32 and 36 is prerequisite to recognition by UBE2D3 leading to polyubiquitination and subsequent degradation.
    Sumoylated; sumoylation requires the presence of the nuclear import signal. Sumoylation blocks ubiquitination and proteasome-mediated degradation of the protein thereby increasing the protein stability.
    Monoubiquitinated at Lys-21 and/or Lys-22 by UBE2D3. Ubiquitin chain elongation is then performed by CDC34 in cooperation with the SCF(FBXW11) E3 ligase complex, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. The resulting polyubiquitination leads to protein degradation. Also ubiquitinated by SCF(BTRC) following stimulus-dependent phosphorylation at Ser-32 and Ser-36.
    Deubiquitinated by porcine reproductive and respiratory syndrome virus Nsp2 protein, which thereby interferes with NFKBIA degradation and impairs subsequent NF-kappa-B activation.
  • 細胞内局在Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export.
  • Information by UniProt
  • 参照データベース
  • 別名
    • I kappa B alpha antibody
    • I-kappa-B-alpha antibody
    • IkappaBalpha antibody
    • IkB-alpha antibody
    • IKBA antibody
    • IKBA_HUMAN antibody
    • IKBalpha antibody
    • MAD 3 antibody
    • MAD3 antibody
    • Major histocompatibility complex enhancer-binding protein MAD3 antibody
    • NF kappa B inhibitor alpha antibody
    • NF-kappa-B inhibitor alpha antibody
    • NFKBI antibody
    • NFKBIA antibody
    • Nuclear factor of kappa light chain gene enhancer in B cells antibody
    • Nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor alpha antibody
    see all

Anti-IKB alpha antibody [EP697] 画像

  • All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/1000 dilution

    Lane 1 : Jurkat (human acute T cell leukemia) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 36 kDa
    Observed band size : 39 kDa (why is the actual band size different from the predicted?)
    Blocking/Diluting buffer 5% NFDM /TBST
  • Immunohistochemical analysis of paraffin-embedded human colon tissue sections labelling IKB alpha with purified ab76429 at dilution of 1/500. Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunocytochemistry/Immunofluorescence staining of HeLa cells labelling IKB alpha with purified ab76429 at a working dilution of 1/250. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. DAPI was used as nuclear counterstain. The cells were fixed in 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, rabbit primary antibody was used followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120). For negative control 2, ab7291 (mouse anti-tubulin) was used followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077).

  • Overlay histogram showing 4% paraformaldehyde fixed Jurkat (human acute T cell leukemia) cells labelling IKB alpha with purified ab76429 at dilution of 1/250. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black line). The blue line shows cells without incubation with primary antibody and secondary antibody. 

  • All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/10000 dilution

    Lane 1 : RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 36 kDa
    Observed band size : 39 kDa (why is the actual band size different from the predicted?)
    Blocking/Diluting buffer 5% NFDM /TBST
  • Immunohistochemical analysis of Paraffin-embedded human colon tissue section labelling IKB alpha with ab76429 at 1/100 dilution. 

  • All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/1000 dilution

    Lane 1 : HeLa cell lysates
    Lane 2 : Jurkat cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    goat anti-rabbit HRP at 1/1000 dilution

    Predicted band size : 36 kDa
    Observed band size : 36 kDa
  • Overlay histogram showing HeLa cells stained with ab76429 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76429, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Anti-IKB alpha antibody [EP697] (ab76429) 使用論文

This product has been referenced in:
  • Zhao Z  et al. Identification of a NFKBIA polymorphism associated with lower NFKBIA protein levels and poor survival outcomes in patients with glioblastoma multiforme. Int J Mol Med 34:1233-40 (2014). IHC ; Human . Read more (PubMed: 25215581) »

See 1 Publication for this product

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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...

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