使用敲除细胞株进行验证RabMAb

Anti-IKK alpha抗体[Y463] (ab32041)

概述

性能

应用

Our Abpromise guarantee covers the use of ab32041 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 1/10000 - 1/50000. Detects a band of approximately 88 kDa (predicted molecular weight: 85 kDa).
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/50.

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

IP 1/50.
ICC/IF 1/500.

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. As part of the non-canonical pathway of NF-kappa-B activation, 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. Also phosphorylates NCOA3. Phosphorylates 'Ser-10' of histone H3 at NF-kappa-B-regulated promoters during inflammatory responses triggered by cytokines.
  • 组织特异性Widely expressed.
  • 疾病相关Defects in CHUK are the cause of cocoon syndrome (COCOS) [MIM:613630]; also known as fetal encasement syndrome. COCOS is a lethal syndrome characterized by multiple fetal malformations including defective face and seemingly absent limbs, which are bound to the trunk and encased under the skin.
  • 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Phosphorylated by MAP3K14/NIK, AKT and to a lesser extent by MEKK1, and dephosphorylated by PP2A. Autophosphorylated.
    Acetylation of Thr-179 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B signaling pathway.
  • 细胞定位Cytoplasm. Nucleus. Shuttles between the cytoplasm and the nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • chuk antibody
    • CHUK1 antibody
    • Conserved Helix Loop Helix Ubiquitous Kinase antibody
    • Conserved helix loop ubiquitous kinase antibody
    • Conserved helix-loop-helix ubiquitous kinase antibody
    • I Kappa B Kinase 1 antibody
    • I Kappa B Kinase Alpha antibody
    • I-kappa-B kinase 1 antibody
    • I-kappa-B kinase alpha antibody
    • IkappaB kinase antibody
    • IkB kinase alpha subunit antibody
    • IkBKA antibody
    • IKK 1 antibody
    • IKK A antibody
    • IKK a kinase antibody
    • IKK-A antibody
    • IKK-alpha antibody
    • IKK1 antibody
    • IKKA antibody
    • IKKA_HUMAN antibody
    • Inhibitor Of Kappa Light Polypeptide Gene Enhancer In B Cells antibody
    • Inhibitor Of Nuclear Factor Kappa B Kinase Alpha Subunit antibody
    • Inhibitor of nuclear factor kappa-B kinase subunit alpha antibody
    • NFKBIKA antibody
    • Nuclear Factor Kappa B Inhibitor Kinase Alpha antibody
    • Nuclear factor NF kappa B inhibitor kinase alpha antibody
    • Nuclear factor NF-kappa-B inhibitor kinase alpha antibody
    • Nuclear factor NFkappaB inhibitor kinase alpha antibody
    • Nuclear Factor Of Kappa Light Chain Gene Enhancer In B Cells Inhibitor antibody
    • TCF-16 antibody
    • TCF16 antibody
    • Transcription factor 16 antibody
    see all

Anti-IKK alpha antibody [Y463] 图像



  • Predicted band size : 85 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: IKK alpha knockout HAP1 cell lysate (20 µg)
    Lanes 1 and 2: Merged signal (red and green). Green - ab32041 observed at 84 kDa. Red - loading control, ab8226, observed at 42 kDa.

    ab32041 was shown to specifically react with IKK alpha when IKK alpha knockout samples were used. Wild-type and IKK alpha knockout samples were subjected to SDS-PAGE. ab32041 and ab8226 (loading control to beta actin) were diluted 1/10 000 and 1/1000 respectively and incubated overnight at 4°C. 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.

  • ab32041 staining IKK alpha in wild-type HAP1 cells (top panel) and IKK alpha 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 ab32041 at 1/500 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.

  • Anti-IKK alpha antibody [Y463] (ab32041) at 1/50000 dilution + Daudi cell lysate

    Predicted band size : 85 kDa
    Observed band size : 88 kDa (why is the actual band size different from the predicted?)
  • Ab32041, at a dilution of 1/50, staining IKK alpha in paraffin embedded human stomach carcinoma by Immunohistochemisty.
  • Overlay histogram showing HeLa cells stained with ab32041 (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 (ab32041, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 85 kDa


    Exposure time : 10 seconds

Anti-IKK alpha antibody [Y463] (ab32041)参考文献

This product has been referenced in:
  • Liu Z  et al. Mark4 promotes oxidative stress and inflammation via binding to PPAR? and activating NF-?B pathway in mice adipocytes. Sci Rep 6:21382 (2016). WB ; Mouse . Read more (PubMed: 26888669) »
  • Jung JU  et al. NIK/MAP3K14 Regulates Mitochondrial Dynamics and Trafficking to Promote Cell Invasion. Curr Biol N/A:N/A (2016). Human . Read more (PubMed: 27889261) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Mouse Cell lysate - whole cell (Raw 264.7)
Total protein in input 100 µg
Specification Raw 264.7
Treatment 2 hrs LPS
Immuno-precipitation step Protein G
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Abcam user community

Verified customer

提交于 Jul 31 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Raw 264.7)
Loading amount 50 µg
Specification Raw 264.7
Treatment 2 hrs LPS
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Abcam user community

Verified customer

提交于 Jun 08 2012

Application Western blot
Sample Guinea pig Tissue lysate - whole (myometrium pregnant and non-pregnant)
Loading amount 30 µg
Specification myometrium pregnant and non-pregnant
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Dec 14 2011

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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Thank you for your enquiry. Unfortunately, the blocking peptide for this product is not available. I am sorry I can't be of much help in this occasion but if there is anything else that I can help you with, please do not hesitate to contact me....

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Epithelial, Neutrophils, monocytes)
Loading amount 500000 cells
Specification Epithelial, Neutrophils, monocytes
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

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (skin)
Specification skin
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6%
Username

Abcam user community

Verified customer

提交于 Apr 24 2007

Thank you for your enquiry. The concentration for the current lot is 0.25mg/ml. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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