Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)

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Western blot - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
  • Flow Cytometry - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
  • Western blot - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
  • Immunocytochemistry/ Immunofluorescence - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
  • OI-RD Scanning - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
  • Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6043] to IKK beta - BSA and Azide free
  • Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-IKK beta antibody [EPR6043] - BSA and Azide free
    See all IKK beta primary antibodies

  • Description

    Rabbit monoclonal [EPR6043] to IKK beta - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human IKK beta aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: O14920

  • General notes

    Ab171364 is the carrier-free version of ab124957. 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.

    ab171364 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.

    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 ab171364 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
WB Use at an assay dependent concentration. Detects a band of approximately 85 kDa (predicted molecular weight: 87 kDa).

Can be blocked with ab154148
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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

 
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

(Heat to 98°C, allow to cool for 10-20 minutes)
  • Application notes
    Is unsuitable for IP.

    • 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

      see all

    • 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

    • Western blot - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      Western blot - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)

      This WB data was generated using the same anti-IKK beta antibody clone [EPR6043] in a different buffer formulation (cat# ab124957).

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: IKK beta knockout HAP1 cell lysate (20 µg) 
      Lanes 1 and 2: Merged signal (red and green). Green - ab124957 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab124957 was shown to specifically react with IKK beta when IKK beta knockout samples were used. Wild-type and IKK beta knockout samples were subjected to SDS-PAGE. ab124957 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/2000 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 1/10,000 dilution for 1 h at room temperature before imaging.

    • Flow Cytometry - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      Flow Cytometry - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)

      Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IKK beta with unpurified ab124957 at 1/20 dilution (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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124957).

    • Western blot - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      Western blot - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      All lanes : Anti-IKK beta antibody [EPR6043] (ab124957) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : IKBKB knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 87 kDa
      Observed band size: 90 kDa
      why is the actual band size different from the predicted?



      This data was developed using the same antibody clone in a different buffer formulation (ab124957).

      Lanes 1- 2: Merged signal (red and green). Green - ab124957 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab124957 was shown to react with IKK beta in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264847 (knockout cell lysate ab257228) lane below 90kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and IKBKB knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab124957 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)

      ab124957, at 1/100 dilution staining IKK beta in paraffin-embedded Human cervix carcinoma tissue, by Immunohistochemistry.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124957).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)

      ab124957, at 1/100 dilution staining IKK beta in paraffin-embedded Mouse kidney tissue, by Immunohistochemistry.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124957).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      Immunocytochemistry/ Immunofluorescence - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      ab124957, at 1/50 dilution, staining IKK beta in HeLa cells by Immunofluorescence.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124957).

    • OI-RD Scanning - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      OI-RD Scanning - Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124957).

    • Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)
      Anti-IKK beta antibody [EPR6043] - BSA and Azide free (ab171364)

    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 ab171364? Please let us know so that we can cite the reference in this datasheet.

    ab171364 has not yet been referenced specifically in any publications.

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