Recombinant Anti-NAK/TBK1 antibody [EPR2867(2)-19] (ab109735)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2867(2)-19] to NAK/TBK1
- Suitable for: WB, IHC-P, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-NAK/TBK1 antibody [EPR2867(2)-19]
See all NAK/TBK1 primary antibodies -
Description
Rabbit monoclonal [EPR2867(2)-19] to NAK/TBK1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IPmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human
Predicted to work with: Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- HeLa, U937, HepG2, and 293T cell lysates, Human testis tissue, HeLa cells
-
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2867(2)-19 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109735 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 | (1) |
1/1000 - 1/10000. Detects a band of approximately 84 kDa (predicted molecular weight: 84 kDa).
For unpurified use at 1/1000 - 1/2000. |
IHC-P |
1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/400. |
|
IP |
1/30.
|
Notes |
---|
WB
1/1000 - 1/10000. Detects a band of approximately 84 kDa (predicted molecular weight: 84 kDa). For unpurified use at 1/1000 - 1/2000. |
IHC-P
1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/400. |
IP
1/30. |
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
- Entrez Gene: 29110 Human
- Entrez Gene: 299827 Rat
- Omim: 604834 Human
- SwissProt: Q9UHD2 Human
- Unigene: 505874 Human
-
Alternative names
- EC 2.7.11.1 antibody
- FLJ11330 antibody
- FTDALS4 antibody
see all
Images
-
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.
-
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, 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 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.
-
Immunohistochemical analysis of paraffin-embedded human testis tissue using ab109735 at a dilution of 1/250.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
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 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.
-
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 (ab172730) instead of ab109735 in HeLa whole cell lysate.
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 -
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 : Anti-NAK/TBK1 antibody [EPR2867(2)-19] (ab109735) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : U937 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : 293T cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 84 kDa
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (17)
ab109735 has been referenced in 17 publications.
- Alshafie W et al. Identification of highly specific antibodies for Serine/threonine-protein kinase TBK1 for use in immunoblot, immunoprecipitation and immunofluorescence. F1000Res 11:977 (2022). PubMed: 36415206
- Tu Y et al. MITA oligomerization upon viral infection is dependent on its N-glycosylation mediated by DDOST. PLoS Pathog 18:e1010989 (2022). PubMed: 36449507
- Weinreich M et al. Neuropathological characterization of a novel TANK binding kinase (TBK1) gene loss of function mutation associated with amyotrophic lateral sclerosis. Neuropathol Appl Neurobiol 46:279-291 (2020). PubMed: 31498468
- Zou HM et al. Human Cytomegalovirus Protein UL94 Targets MITA to Evade the Antiviral Immune Response. J Virol 94:N/A (2020). PubMed: 32238587
- Ran Y et al. YIPF5 Is Essential for Innate Immunity to DNA Virus and Facilitates COPII-Dependent STING Trafficking. J Immunol 203:1560-1570 (2019). PubMed: 31391232