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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)

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Western blot - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
  • Immunocytochemistry - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
  • Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
  • Western blot - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
  • Immunoprecipitation - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
  • Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR464] to IKK alpha - BSA and Azide free
  • Suitable for: ICC, IHC-P, IP, Flow Cyt (Intra), WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-IKK alpha antibody [EPR464] - BSA and Azide free
    See all IKK alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR464] to IKK alpha - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, IHC-P, IP, Flow Cyt (Intra), WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Jurkat, HeLa, HepG2 and Daudi cell lysates; ICC: HeLa cells; IHC: Human breast tissue IP: Jurkat cell lysate. Flow Cyt: HeLa cells.

  • General notes

    ab171362 is the carrier-free version of ab109749.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab171362 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
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 85 kDa.
Notes
ICC
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 85 kDa.

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

    • Western blot - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      Western blot - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)

      This data was developed using ab109749, the same antibody clone in a different buffer formulation.

      Lanes 1, 3, and 5:ild-type HAP1 cell lysate (20 µg)

      Lanes 2, 4, and 6: IKK alpha knockout HAP1 cell lysate (20 µg)

      Lanes 1 and 2: Green signal from target - ab109749 observed at 84 kDa

      Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa

      Lanes 5 and 6: Merged (red and green) signal

      ab109749 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. ab109749 and ab8226 (loading control to beta actin) were both diluted at 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      Immunocytochemistry - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      This data was developed using ab109749, the same antibody clone in a different buffer formulation.Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling IKK alpha with purified ab109749 at 1/250 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor®488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
    • Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)

      This data was developed using ab109749, the same antibody clone in a different buffer formulation. 

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IKK alpha with unpurified ab109749 at 1/50 dilutio n (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

       

       

    • Western blot - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      Western blot - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      All lanes : Anti-IKK alpha antibody [EPR464] (ab109749) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : Daudi cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 85 kDa



      This data was developed using ab109749, the same antibody clone in a different buffer formulation.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      This data was developed using ab109749, the same antibody clone in a different buffer formulation.ab109749, at 1/100 dilution, staining IKK alpha in paraffin-embedded Human breast tissue by Immunohistochemistry. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
    • Immunoprecipitation - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      Immunoprecipitation - Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)

      This data was developed using ab109749, the same antibody clone in a different buffer formulation.

      IKK alpha was immunoprecipitated from 0.35 mg Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 µg with ab109749 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 µg

      Lane 2: abab109749 IP in Jurkat whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab109749 in Jurkat whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    • Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)
      Anti-IKK alpha antibody [EPR464] - BSA and Azide free (ab171362)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab171362? Please let us know so that we can cite the reference in this datasheet.

    ab171362 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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