Validated using a knockout cell line
Recombinant
RabMAb

Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)

Overview

  • Product name
    Anti-IKK alpha antibody [Y463] - BSA and Azide free
    See all IKK alpha primary antibodies
  • Description
    Rabbit monoclonal [Y463] to IKK alpha - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-P, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control
    • WB: Daudi cell lysate.
  • General notes

    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.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab169743 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 at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 88 kDa (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

  • Overlay histogram showing HAP1 wildtype (green line) and HAP1-CHUK knockout cells (red line) stained with ab32041. 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 (ab32041, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C.

    A rabbit IgG isotype control antibody  (ab172730) 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 with80% methanol (5 min), , permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32041).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32041).

  • Ab32041, at a dilution of 1/50, staining IKK alpha in paraffin embedded human stomach carcinoma by Immunohistochemisty.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32041).

  • 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 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32041).

References

This product has been referenced in:
See 1 Publication for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab169743.
Please use the links above to contact us or submit feedback about this product.

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

Sign up