Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-IKK alpha antibody [Y463] (Phycoerythrin) (ab210716)

Overview

  • Product name
    Anti-IKK alpha antibody [Y463] (Phycoerythrin)
    See all IKK alpha primary antibodies
  • Description
    Rabbit monoclonal [Y463] to IKK alpha (Phycoerythrin)
  • Host species
    Rabbit
  • Conjugation
    Phycoerythrin. Ex: 488nm, Em: 575nm
  • Tested applications
    Suitable for: ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human IKK alpha aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: O15111

  • Positive control
    • Flow Cyt: HeLa cells, HAP1-WT cells. ICC/IF: HeLa cells
  • General notes

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

Properties

Applications

Our Abpromise guarantee covers the use of ab210716 in the following tested applications.

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

Application Abreviews Notes
ICC/IF 1/50.

This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)

Flow Cyt 1/500.

Target

  • Function
    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.
  • Tissue specificity
    Widely expressed.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Cytoplasm. Nucleus. Shuttles between the cytoplasm and the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • Overlay histogram showing HAP1 wildtype (green line) and HAP1-CHUK knockout cells (red line) stained with ab210716. The cells were fixed with 4% formaldehyde (10 min) , and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab210716, 0.5µg/ml dilution) for 30 min at 22°C.

    A rabbit monoclonal IgG isotype control antibody  (ab209478) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CHUK  knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

    This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min) permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

  • Overlay histogram showing HeLa cells stained with ab210716 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol for 30 min at -20°C. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab210716, 1/500 dilution) for 30 min at 22°C.

    Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50mW Yellow-Green laser (561nm) and 586/15 bandpass filter.

  • ab210716 staining IKK alpha in HeLa cells. The cells were fixed with 100% methanol (5min), 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 overnight at +4°C with ab210716 at 1/50 dilution (pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).

References

ab210716 has not yet been referenced specifically in any publications.

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