Validated using a knockout cell lineRecombinantRabMAb

Recombinant Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)

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Flow Cytometry - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
  • Flow Cytometry - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
  • Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Alexa Fluor® 488 Rabbit monoclonal [Y463] to IKK alpha
  • Suitable for: ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm

Overview

  • Product name

    Alexa Fluor® 488 Anti-IKK alpha antibody [Y463]
    See all IKK alpha primary antibodies

  • Description

    Alexa Fluor® 488 Rabbit monoclonal [Y463] to IKK alpha

  • Host species

    Rabbit

  • Conjugation

    Alexa Fluor® 488. Ex: 495nm, Em: 519nm

  • 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.

  • Positive control

    • ICC/IF: HeLa and wildtype HAP1 cells. Flow Cyt: HeLa cells.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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab200412 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 1/100 - 1/400.
Flow Cyt 1/50.

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

    • Flow Cytometry - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
      Flow Cytometry - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)

      Overlay histogram showing HAP1 wildtype (green line) and HAP1-CHUK knockout cells (red line) stained with ab200412. 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 (ab200412, 0.1µg/ml dilution) for 30 min at 22°C.

      A rabbit monoclonal IgG isotype control antibody  (ab199091) 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 Blue laser (488nm) and 530/30 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.

    • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
      Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)

      ab200412 staining IKKα in wild-type HAP1 cells (top panel) and IKKα knockout HAP1 cells (bottom panel). 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 with ab200412 at 1/400 dilution and ab7291 at 1μg/ml dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo colour red). Nuclear DNA was labelled in blue with DAPI.

    • Flow Cytometry - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
      Flow Cytometry - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
      Overlay histogram showing HeLa cells stained with ab200412 (red line). The cells were fixed with 4% formaldehyde (10 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 (ab200412, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG [EPR25A] Alexa Fluor® 488 (ab199091) 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 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
      Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)

      ab200412 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 ab200412 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

    • Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
      Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)

    References (0)

    Publishing research using ab200412? Please let us know so that we can cite the reference in this datasheet.

    ab200412 has not yet been referenced specifically in any publications.

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