Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-IKK beta antibody [Y466] - BSA and Azide free (ab171363)

Overview

  • Product name

    Anti-IKK beta antibody [Y466] - BSA and Azide free
    See all IKK beta primary antibodies
  • Description

    Rabbit monoclonal [Y466] to IKK beta - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human IKK beta aa 650-750 (C terminal). The exact sequence is proprietary.
    Database link: O14920

  • Positive control

    • WB: Wild-type HAP1 cell lysate.
  • General notes

    Ab171363 is the carrier-free version of ab32135. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab171363 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab171363 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
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 87 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. Also phosphorylates NCOA3.
  • Tissue specificity

    Highly expressed in heart, placenta, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis and peripheral blood.
  • 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

    Upon cytokine stimulation, phosphorylated on Ser-177 and Ser-181 by MEKK1 and/or MAP3K14/NIK; which enhances activity. Once activated, autophosphorylates on the C-terminal serine cluster; which decreases activity and prevents prolonged activation of the inflammatory response.
    Acetylation of Thr-180 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B pathway.
    Ubiquitinated. Monoubiquitination involves TRIM21 that leads to inhibition of Tax-induced NF-kappa-B signaling. According to PubMed:19675099, 'Ser-163' does not serve as a monoubiquitination site. According to PubMed:16267042, ubiquitination on 'Ser-163' modulates phosphorylation on C-terminal serine residues. Monoubiquitination by TRIM21 is dirupted by Yersinia yopJ.
  • Cellular localization

    Cytoplasm. Membrane raft. Colocalized with DPP4 in membrane rafts.
  • Information by UniProt
  • Database links

  • Alternative names

    • I kappa B kinase 2 antibody
    • I kappa B kinase beta antibody
    • I-kappa-B kinase 2 antibody
    • I-kappa-B-kinase beta antibody
    • IkBKB antibody
    • IKK beta antibody
    • IKK-B antibody
    • IKK-beta antibody
    • IKK2 antibody
    • IKKB antibody
    • IKKB_HUMAN antibody
    • IMD15 antibody
    • Inhibitor of kappa light polypeptide gene enhancer in B cells, kinase beta antibody
    • Inhibitor of nuclear factor kappa-B kinase subunit beta antibody
    • NFKBIKB antibody
    • Nuclear factor NF-kappa-B inhibitor kinase beta antibody
    see all

Images

  • ab32135 (purified) at 1:50 dilution (2μg) immunoprecipitating IKK beta in SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate.

    Lane 1 (input): SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab32135 & SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32135 in SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:10,000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32135).

  • Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 4 and 6: IKK beta knockout HAP1 cell lysate (20 µg)
    Lanes 1 and 2: Green signal from target – ab32135 observed at 87 kDa
    Lanes 3 and 4: Red signal from loading control – ab8226 observed at 42 kDa
    Lanes 5 and 6: Merged (red and green) signal

    Unpurified ab32135 was shown to react with IKK beta when IKK beta knockout samples were used, along with additional cross-reactive bands. Wild-type and IKK beta knockout samples were subjected to SDS-PAGE. ab32135 and ab8226 (loading control to beta actin) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4ºC. 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 1/10 000 dilution for 1 h at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32135).

References

ab171363 has not yet been referenced specifically in any publications.

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