Rabbit Recombinant Monoclonal NAK/TBK1 antibody. Suitable for IP, WB, IHC-P and reacts with Human samples. Cited in 20 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | Flow Cyt | WB | IHC-P | |
---|---|---|---|---|
Human | Tested | Not recommended | Tested | Tested |
Rat | Predicted | Not recommended | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes For unpurified use at 1/50. Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 - 1/10000 | Notes For unpurified use at 1/1000 - 1/2000. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/800 | Notes For unpurified use at 1/400. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
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Serine/threonine kinase that plays an essential role in regulating inflammatory responses to foreign agents (PubMed:10581243, PubMed:11839743, PubMed:12692549, PubMed:12702806, PubMed:14703513, PubMed:15367631, PubMed:15485837, PubMed:18583960, PubMed:21138416, PubMed:23453971, PubMed:23453972, PubMed:23746807, PubMed:25636800, PubMed:26611359, PubMed:32404352, PubMed:34363755). Following activation of toll-like receptors by viral or bacterial components, associates with TRAF3 and TANK and phosphorylates interferon regulatory factors (IRFs) IRF3 and IRF7 as well as DDX3X (PubMed:12692549, PubMed:12702806, PubMed:14703513, PubMed:15367631, PubMed:18583960, PubMed:25636800). This activity allows subsequent homodimerization and nuclear translocation of the IRFs leading to transcriptional activation of pro-inflammatory and antiviral genes including IFNA and IFNB (PubMed:12702806, PubMed:15367631, PubMed:25636800, PubMed:32972995). In order to establish such an antiviral state, TBK1 form several different complexes whose composition depends on the type of cell and cellular stimuli (PubMed:23453971, PubMed:23453972, PubMed:23746807). Plays a key role in IRF3 activation: acts by first phosphorylating innate adapter proteins MAVS, STING1 and TICAM1 on their pLxIS motif, leading to recruitment of IRF3, thereby licensing IRF3 for phosphorylation by TBK1 (PubMed:25636800, PubMed:30842653). Phosphorylated IRF3 dissociates from the adapter proteins, dimerizes, and then enters the nucleus to induce expression of interferons (PubMed:25636800). Thus, several scaffolding molecules including FADD, TRADD, MAVS, AZI2, TANK or TBKBP1/SINTBAD can be recruited to the TBK1-containing-complexes (PubMed:21931631). Under particular conditions, functions as a NF-kappa-B effector by phosphorylating NF-kappa-B inhibitor alpha/NFKBIA, IKBKB or RELA to translocate NF-Kappa-B to the nucleus (PubMed:10783893, PubMed:15489227). Restricts bacterial proliferation by phosphorylating the autophagy receptor OPTN/Optineurin on 'Ser-177', thus enhancing LC3 binding affinity and antibacterial autophagy (PubMed:21617041). Phosphorylates SMCR8 component of the C9orf72-SMCR8 complex, promoting autophagosome maturation (PubMed:27103069). Phosphorylates ATG8 proteins MAP1LC3C and GABARAPL2, thereby preventing their delipidation and premature removal from nascent autophagosomes (PubMed:31709703). Phosphorylates and activates AKT1 (PubMed:21464307). Seems to play a role in energy balance regulation by sustaining a state of chronic, low-grade inflammation in obesity, wich leads to a negative impact on insulin sensitivity (By similarity). Attenuates retroviral budding by phosphorylating the endosomal sorting complex required for transport-I (ESCRT-I) subunit VPS37C (PubMed:21270402). Phosphorylates Borna disease virus (BDV) P protein (PubMed:16155125). Plays an essential role in the TLR3- and IFN-dependent control of herpes virus HSV-1 and HSV-2 infections in the central nervous system (PubMed:22851595). Acts both as a positive and negative regulator of the mTORC1 complex, depending on the context: activates mTORC1 in response to growth factors by catalyzing phosphorylation of MTOR, while it limits the mTORC1 complex by promoting phosphorylation of RPTOR (PubMed:29150432, PubMed:31530866). Involved in the regulation of TNF-induced RIPK1-mediated cell death, probably acting via CYLD phosphorylation that in turn controls RIPK1 ubiquitination status (PubMed:34363755). Participates also in the differentiation of T follicular regulatory cells together with the receptor ICOS (PubMed:27135603).
NAK, NAK, TBK1, Serine/threonine-protein kinase TBK1, NF-kappa-B-activating kinase, T2K, TANK-binding kinase 1
Rabbit Recombinant Monoclonal NAK/TBK1 antibody. Suitable for IP, WB, IHC-P and reacts with Human samples. Cited in 20 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR2867(2)-19
Affinity purification Protein A
This antibody may have weak cross-reactivity with IKBKE (IKKε).
Blue Ice
+4°C
-20°C
Stable for 12 months at -20°C
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 monoclonal antibody, which offers several advantages including:
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This supplementary information is collated from multiple sources and compiled automatically.
The NAK/TBK1 also known as TANK-binding kinase 1 is a fundamental protein kinase in various cellular processes. It weighs approximately 84 kDa and is expressed in many tissues including lymphoid organs and the brain. NAK acts as a serine/threonine-protein kinase which transfers a phosphate group to specific substrates modulating their function and activity. Phosphorylation by TBK1 initiates several signaling events making it an important regulator in immune responses and inflammatory signaling pathways.
This kinase plays a significant role in antiviral defense and autophagy. NAK/TBK1 is also involved in innate immune signaling and cell survival. TBK1 forms part of a larger signaling complex interacting with adaptors like TRAF3 and TANK which are important for its proper localization and function. Phospho-NAK its phosphorylated form can activate other molecules driving various downstream immune and stress responses.
The protein is integral to the type I interferon signaling pathway and NF-kB pathway. NAK/TBK1 interacts closely with IRF3 a transcription factor important for antiviral activity and plays a role alongside other proteins like IKKε. These interactions permit activation of genes necessary for defense mechanisms against viral pathogens and control of inflammation. In the NF-kB pathway TBK1 helps regulate cellular responses to stress and inflammatory signals.
NAK/TBK1 has associations with amyotrophic lateral sclerosis (ALS) and various cancers. Research shows connections between TBK1 mutations and ALS indicating its importance in neuronal function and health. Additionally aberrations in TBK1 activity contribute to oncogenesis as it can affect cell proliferation and survival. The protein's relation with signaling proteins such as p62/SQSTM1 highlights its diverse role in disease mechanisms making it a potential therapeutic target.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Immunoprecipitation of TBK1 in HeLa cells. Lysates were prepared and immunoprecipitation was performed using 1.0 μg of ab109735 pre-coupled to prot.A-Sepharose beads. Samples were washed and processed for western blot with Anti-NAK/TBK1 antibody [108A429] ab12116 at 1 in 2000. This data was kindly provided by the YCharOS Inc., an open science company with the mission of characterizing every commercially available antibody reagent. Abcam are working with YCharOS to support their mission of antibody characterisation using knock out cell lines.
All lanes: Immunoprecipitation - Anti-NAK/TBK1 antibody [EPR2867(2)-19] (ab109735)
Predicted band size: 84 kDa
ab109735 was shown to react with TBK1 in wild-type U2OSn cells in Western blot with loss of signal observed in a TBK1 knockout cell line. Wild-type U2OSn and TBK1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab109735 overnight at 4 °C at a 1/10000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 1/5000 before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-NAK/TBK1 antibody [EPR2867(2)-19] (ab109735)
Predicted band size: 84 kDa
Purified ab109735 at 1/30 dilution (2μg) immunoprecipitating NAK/TBK1 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+): ab109735 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109735 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 84 kDa
All lanes: Immunoprecipitation - Anti-NAK/TBK1 antibody [EPR2867(2)-19] (ab109735)
Predicted band size: 61 kDa, 84 kDa, 94 kDa
Observed band size: 85 kDa
Lane 1: Wild-type HAP1 cell lysate (20 μg)
Lane 2: NAK/TBK1 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 - ab109735 observed at 90 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab109735 was shown to recognize NAK/TBK1 when NAK/TBK1 knockout samples were used, along with additional cross-reactive bands. Wild-type and NAK/TBK1 knockout samples were subjected to SDS-PAGE. ab109735 and Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-NAK/TBK1 antibody [EPR2867(2)-19] (ab109735)
Predicted band size: 84 kDa
ab109735 staining NAK/TBK1 in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/800). An undiluted HRP-conjugated anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin.
All lanes: Western blot - Anti-NAK/TBK1 antibody [EPR2867(2)-19] (ab109735) at 1/1000 dilution
Lane 1: HeLa cell lysate at 10 µg
Lane 2: U937 cell lysate at 10 µg
Lane 3: HepG2 cell lysate at 10 µg
Lane 4: 293T cell lysate at 10 µg
Predicted band size: 84 kDa
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