Validated using a knockout cell line

Anti-NAK/TBK1 antibody [108A429] (ab12116)

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Key features and details

  • Mouse monoclonal [108A429] to NAK/TBK1
  • Suitable for: Flow Cyt, WB, IP
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-NAK/TBK1 antibody [108A429]
    See all NAK/TBK1 primary antibodies

  • Description

    Mouse monoclonal [108A429] to NAK/TBK1

  • Host species

    Mouse

  • Specificity

    ab12116 recognises human NAK.

  • Tested applications

    Suitable for: Flow Cyt, WB, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is considered to be commercially sensitive.

  • Positive control

    • 293 cell extracts transfected with human TBK1 cDNA.

Properties

Applications

Our Abpromise guarantee covers the use of ab12116 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 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 87.5 kDa).
IP Use at an assay dependent concentration.

Target

  • Function

    Serine/threonine protein involved in the signaling cascade converging to the activation of the transcription factor NF-kappa-B. May function as an IKK kinase, playing an essential role in the transcription of a subset of TNF-alpha-induced genes. Also mediates production of RANTES/CCL5 and interferon-beta/IFNB1. Has a pivotal role in the innate immune response. Phosphorylates Borna disease virus (BDV) P protein. Phosphorylates and activates IRF3 and IRF7 and allows their nuclear localization. This leads to production of alpha/beta interferons and the development of a cellular antiviral state. It also seems to be a central factor in the induction of the antiviral interferon response. Inhibition of its interaction with IRF3, due to HCV NS3 binding or BDV P protein seems to be one mechanism of inhibition of the innate immune responses of hepatitis C virus (HCV) infection or Borna disease virus infection respectively.

  • Tissue specificity

    Ubiquitous with higher expression in testis.

  • Sequence similarities

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
    Contains 1 protein kinase domain.

  • Cellular localization

    Cytoplasm.
  • Information by UniProt

  • Database links

    • Alternative names

      • EC 2.7.11.1 antibody
      • FLJ11330 antibody
      • FTDALS4 antibody
      • NAK antibody
      • NF kappa B activating kinase antibody
      • NF kB activating kinase antibody
      • NF-kappa-B-activating kinase antibody
      • Serine/threonine protein kinase TBK 1 antibody
      • Serine/threonine protein kinase TBK1 antibody
      • Serine/threonine-protein kinase TBK1 antibody
      • T2K antibody
      • TANK binding kinase 1 antibody
      • TANK-binding kinase 1 antibody
      • TBK 1 antibody
      • Tbk1 antibody
      • TBK1_HUMAN antibody
      see all

    Images

    • Western blot - Anti-NAK/TBK1 antibody [108A429] (ab12116)
      Western blot - Anti-NAK/TBK1 antibody [108A429] (ab12116)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: NAK knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: HepG2 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab12116 observed at 90 kDa. Red - loading control, ab181602, observed at 37 kDa.
      ab12116 was shown to recognize NAK when NAK knockout samples were used, along with additional cross-reactive bands. Wild-type and NAK knockout samples were subjected to SDS-PAGE. ab12116 and ab181602 (loading control to GAPDH) were diluted at 1 μg/ml and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-NAK/TBK1 antibody [108A429] (ab12116)
      Western blot - Anti-NAK/TBK1 antibody [108A429] (ab12116)
      All lanes : Anti-NAK/TBK1 antibody [108A429] (ab12116) at 2 µg/ml

      Lane 1 : 293 cells
      Lane 2 : 293 cells transfected with Human NAK cDNA

      Predicted band size: 87.5 kDa

    • Flow Cytometry - Anti-NAK/TBK1 antibody [108A429] (ab12116)
      Flow Cytometry - Anti-NAK/TBK1 antibody [108A429] (ab12116)
      Overlay histogram showing HeLa cells stained with ab12116 (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. The cells were then incubated with the antibody (ab12116, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

    References

    Publishing research using ab12116? Please let us know so that we can cite the reference in this datasheet.

    ab12116 has been referenced in 5 publications.

    • Ahn J  et al. STING-Dependent Signaling Underlies IL-10 Controlled Inflammatory Colitis. Cell Rep 21:3873-3884 (2017). PubMed: 29281834
    • Lio CW  et al. cGAS-STING Signaling Regulates Initial Innate Control of Cytomegalovirus Infection. J Virol 90:7789-97 (2016). PubMed: 27334590
    • Charoenthongtrakul S  et al. RING finger protein 11 targets TBK1/IKKi kinases to inhibit antiviral signaling. PLoS One 8:e53717 (2013). WB . PubMed: 23308279
    • Gao L  et al. ABIN1 Protein Cooperates with TAX1BP1 and A20 Proteins to Inhibit Antiviral Signaling. J Biol Chem 286:36592-602 (2011). WB, IP . PubMed: 21885437
    • Parvatiyar K  et al. TAX1BP1 and A20 inhibit antiviral signaling by targeting TBK1-IKKi kinases. J Biol Chem 285:14999-5009 (2010). WB, IP ; Human . PubMed: 20304918

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