Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-IKK alpha antibody [Y463] (Alexa Fluor® 647) (ab200414)

Overview

  • Product name
    Anti-IKK alpha antibody [Y463] (Alexa Fluor® 647)
    See all IKK alpha primary antibodies
  • Description
    Rabbit monoclonal [Y463] to IKK alpha (Alexa Fluor® 647)
  • Host species
    Rabbit
  • Conjugation
    Alexa Fluor® 647. Ex: 652nm, Em: 668nm
  • Tested applications
    Suitable for: Flow Cyt, ICC/IFmore 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.

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

    Alternative versions available:
    Anti-IKK alpha antibody [Y463] (ab32041) - Knockout validated

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

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab200414 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
Flow Cyt Use a concentration of 0.1 µg/ml.
ICC/IF 1/100 - 1/200.

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 ab200414. The cells were fixed with 80% methanol (5 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 (ab200414, 0.1µg/ml dilution) for 30 min at 22°C.

    A rabbit monoclonal IgG isotype control antibody  (ab199093) 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 40 mW Red laser (640nm) and 670/14 bandpass filter.

    This antibody can also be used in HAP1 cells fixed with 4% formaldehyde (10 min) permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

  • ab200414 staining IKK alpha in HeLa cells. 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 overnight at +4°C with ab200414 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 2µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

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

  • ab200414 staining IKK alpha in wild-type HAP1 cells (top panel) and IKK alpha knockout HAP1 cells (bottom panel). The cells were fixed with 100% MeOH (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 with ab200414 at 1/200 dilution (shown in red) and ab7291 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

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

References

ab200414 has not yet been referenced specifically in any publications.

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