使用敲除细胞株进行验证RabMAb

Anti-IKK beta抗体[Y466] (ab32135)

概述

  • 产品名称Anti-IKK beta抗体[Y466]
    参阅全部 IKK beta 一抗
  • 描述
    兔单克隆抗体[Y466] to IKK beta
  • 经测试应用适用于: WB, IPmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    A synthetic peptide corresponding to residues near the C-term of human IKK-beta.

  • 阳性对照
    • Daudi cell lysate and human cervical carcinoma.
  • 常规说明

    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.

    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.

    Alternative versions available:

    Anti-IKK beta antibody (HRP) [Y466] (ab194713)

性能

应用

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.

应用 Ab评论 说明
WB 1/1000 - 1/5000. Detects a band of approximately 87 kDa.
IP 1/50.

靶标

  • 功能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.
  • 组织特异性Highly expressed in heart, placenta, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis and peripheral blood.
  • 序列相似性Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
    Contains 1 protein kinase domain.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm. Membrane raft. Colocalized with DPP4 in membrane rafts.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-IKK beta antibody [Y466] 图像

  • 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

    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) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

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

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


  • Exposure time : 3 minutes

Anti-IKK beta antibody [Y466] (ab32135)参考文献

This product has been referenced in:
  • Yu Z  et al. Lys29-linkage of ASK1 by Skp1-Cullin 1-Fbxo21 ubiquitin ligase complex is required for antiviral innate response. Elife 5:N/A (2016). Read more (PubMed: 27063938) »
  • Fan HY  et al. Salvianolic acid A as a multifunctional agent ameliorates doxorubicin-induced nephropathy in rats. Sci Rep 5:12273 (2015). Rat . Read more (PubMed: 26194431) »

See all 7 Publications for this product

Product Wall

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

Ms. kyungjoo seong

Verified customer

提交于 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

提交于 Sep 02 2009

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