corresponding to Human NAK/TBK1 aa 1 to the C-terminus (N terminal).
WB: HepG2, SH-SY5Y, C6, HAP1 and NIH/3T3 cell lysate
IHC-P: Human hepatocellular carcinoma
ICC/IF: MCF7 cells
The mouse and rat recommendation. This antibody may not be suitable for IHC with mouse or rat samples.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
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.
Ubiquitous with higher expression in testis.
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily. Contains 1 protein kinase domain.
Western blot - Anti-NAK antibody [EP611Y] (ab40676)
Predicted band size : 84 kDa
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 - ab40676 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa. ab40676 was shown to specifically react with NAK when NAK knockout samples were used. Wild-type and NAK knockout samples were subjected to SDS-PAGE. ab40676 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10,000 respectively 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.
Western blot - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
All lanes : Anti-NAK/TBK1 antibody [EP611Y] (ab40676) at 1/5000 dilution (purified)
Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate Lane 2 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size : 84 kDa Observed band size : 84 kDa Blocking/Diluting buffer 5% NFDM/TBST
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue sections labeling NAK/TBK1 with purified ab40676 at a dilution of 1/100 (11.5 μg/ml). ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 was used as the secondary anitbody. Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated using EDTA Buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Immunocytochemistry/Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma) cells labeling NAK/TBK1 with purified ab40676 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluo®488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) at 1/200. DAPI (blue) was used as a nuclear counterstain. Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
This product has been referenced in:
Chunfa L et al. The Central Role of IFI204 in IFN-ß Release and Autophagy Activation during Mycobacterium bovis Infection. Front Cell Infect Microbiol7:169 (2017).
Read more (PubMed: 28529930) »
Thurston TL et al. Recruitment of TBK1 to cytosol-invading Salmonella induces WIPI2-dependent antibacterial autophagy. EMBO J35:1779-92 (2016).
Read more (PubMed: 27370208) »