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RabMAb

Anti-IKK beta 抗体 [Y466] (ab32135)

製品の概要

  • 製品名
    Anti-IKK beta antibody [Y466]
    IKK beta 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [Y466] to IKK beta
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: WB, IPmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Synthetic peptide within Human IKK beta aa 650-750 (C terminal). The exact sequence is proprietary.
    Database link: O14920

  • ポジティブ・コントロール
    • WB: Wild-type HAP1 cell lysate. HeLa, NIH/3T3 and Daudi cell lysate. IHC-P: Human cervical carcinoma tissue. IP: SH-SY5Y whole cell lysate.
  • 特記事項

    A trial size is available to purchase for this antibody.

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

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

    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.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32135 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 1/1000 - 1/5000. Detects a band of approximately 87 kDa.

This antibody has weaker binding with mouse than human.

IP 1/50.

ターゲット情報

  • 機能
    Acts as part of the IKK complex in the conventional pathway of NF-kappa-B activation and phosphorylates inhibitors of NF-kappa-B thus leading to the dissociation of the inhibitor/NF-kappa-B complex and ultimately the degradation of the inhibitor. Also phosphorylates NCOA3.
  • 組織特異性
    Highly expressed in heart, placenta, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis and peripheral blood.
  • 配列類似性
    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
    Contains 1 protein kinase domain.
  • 翻訳後修飾
    Upon cytokine stimulation, phosphorylated on Ser-177 and Ser-181 by MEKK1 and/or MAP3K14/NIK; which enhances activity. Once activated, autophosphorylates on the C-terminal serine cluster; which decreases activity and prevents prolonged activation of the inflammatory response.
    Acetylation of Thr-180 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B pathway.
    Ubiquitinated. Monoubiquitination involves TRIM21 that leads to inhibition of Tax-induced NF-kappa-B signaling. According to PubMed:19675099, 'Ser-163' does not serve as a monoubiquitination site. According to PubMed:16267042, ubiquitination on 'Ser-163' modulates phosphorylation on C-terminal serine residues. Monoubiquitination by TRIM21 is dirupted by Yersinia yopJ.
  • 細胞内局在
    Cytoplasm. Membrane raft. Colocalized with DPP4 in membrane rafts.
  • Information by UniProt
  • 参照データベース
  • 別名
    • I kappa B kinase 2 antibody
    • I kappa B kinase beta antibody
    • I-kappa-B kinase 2 antibody
    • I-kappa-B-kinase beta antibody
    • IkBKB antibody
    • IKK beta antibody
    • IKK-B antibody
    • IKK-beta antibody
    • IKK2 antibody
    • IKKB antibody
    • IKKB_HUMAN antibody
    • IMD15 antibody
    • Inhibitor of kappa light polypeptide gene enhancer in B cells, kinase beta antibody
    • Inhibitor of nuclear factor kappa-B kinase subunit beta antibody
    • NFKBIKB antibody
    • Nuclear factor NF-kappa-B inhibitor kinase beta antibody
    see all

画像

  • ab32135 (purified) at 1:50 dilution (2μg) immunoprecipitating IKK beta in SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate.

    Lane 1 (input): SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab32135 & SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32135 in SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate

    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:10,000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 4 and 6: IKK beta knockout HAP1 cell lysate (20 µg)
    Lanes 1 and 2: Green signal from target – ab32135 observed at 87 kDa
    Lanes 3 and 4: Red signal from loading control – ab8226 observed at 42 kDa
    Lanes 5 and 6: Merged (red and green) signal

    Unpurified ab32135 was shown to react with IKK beta when IKK beta knockout samples were used, along with additional cross-reactive bands. Wild-type and IKK beta knockout samples were subjected to SDS-PAGE. ab32135 and ab8226 (loading control to beta actin) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4ºC. 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-IKK beta antibody [Y466] (ab32135) at 1/1000 dilution (purified)

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
    Lane 2 : Mouse lung lysates
    Lane 3 : Rat brain lysates
    Lane 4 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates

    Lysates/proteins at 20 µg per lane.

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

    Observed band size: 87 kDa (why is the actual band size different from the predicted?)



    Blocking and diluting buffer: 5% NFDM/TBST

     

  • Anti-IKK beta antibody [Y466] (ab32135) at 1/2500 dilution (purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

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


    Blocking and diluting buffer: 5% NFDM/TBST

  • Anti-IKK beta antibody [Y466] (ab32135) at 1/5000 dilution (unpurified) + Daudi cell lysate

    Observed band size: 87 kDa (why is the actual band size different from the predicted?)

  • unpurified ab32135

参考文献

This product has been referenced in:
  • Wen H  et al. Recurrent ECSIT mutation encoding V140A triggers hyperinflammation and promotes hemophagocytic syndrome in extranodal NK/T cell lymphoma. Nat Med 24:154-164 (2018). WB . Read more (PubMed: 29291352) »
  • Mettang M  et al. IKK2/NF-?B signaling protects neurons after traumatic brain injury. FASEB J 32:1916-1932 (2018). Read more (PubMed: 29187362) »

See all 13 Publications for this product

レビューと Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Purified protein (Neural stem cells)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
20 µg
Specification
Neural stem cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Kyungjoo Seong

Verified customer

投稿 Jan 21 2013

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
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Epithelial, neutrophil, monocyte)
Loading amount
500000 cells
Specification
Epithelial, neutrophil, monocyte
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Sep 02 2009

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

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