Recombinant Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP611Y] to NAK/TBK1
- Suitable for: ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-NAK/TBK1 antibody [EP611Y]
See all NAK/TBK1 primary antibodies -
Description
Rabbit monoclonal [EP611Y] to NAK/TBK1 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human NAK/TBK1 aa 1-100 (N terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa membrane extract lysate (ab29547), HepG2, SH-SY5Y, C6, HAP1 and NIH/3T3 cell lysate IHC-P: Human hepatocellular carcinoma ICC/IF: MCF7 cells
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General notes
Anti-NAK/TBK1 antibody [EP611Y] (ab40676) may not be suitable for IHC with mouse or rat samples.
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.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP611Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40676 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 |
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ICC/IF |
1/100.
For unpurified use at 1/250 - 1/500. |
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WB | (2) |
1/5000. Detects a band of approximately 84 kDa (predicted molecular weight: 84 kDa).
For unpurified use at 1/1000. |
IHC-P |
1/100. 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 1/50. |
Notes |
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ICC/IF
1/100. For unpurified use at 1/250 - 1/500. |
WB
1/5000. Detects a band of approximately 84 kDa (predicted molecular weight: 84 kDa). For unpurified use at 1/1000. |
IHC-P
1/100. 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 1/50. |
Target
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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
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Database links
- Entrez Gene: 29110 Human
- Entrez Gene: 56480 Mouse
- Entrez Gene: 299827 Rat
- Omim: 604834 Human
- SwissProt: Q9UHD2 Human
- SwissProt: Q9WUN2 Mouse
- Unigene: 505874 Human
- Unigene: 34580 Mouse
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Alternative names
- EC 2.7.11.1 antibody
- FLJ11330 antibody
- FTDALS4 antibody
see all
Images
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ab40676 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 ab40676 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.
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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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
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.
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ab40676 was shown to react with TBK1 in wild-type U2OSn cells in Immunocytochemistry with loss of signal observed in a TBK1 knockout cell line. Wild-type and Knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 5%BSA+5%goat serum (30min). The cells were then incubated with ab40676 at 1/1500 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).
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 : 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
All lanes : 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 -
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.
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.
Datasheets and documents
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SDS download
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Datasheet download
References (86)
ab40676 has been referenced in 86 publications.
- Mukai K et al. Homeostatic regulation of STING by retrograde membrane traffic to the ER. Nat Commun 12:61 (2021). PubMed: 33397928
- Banerjee A et al. Experimental and natural evidence of SARS-CoV-2-infection-induced activation of type I interferon responses. iScience 24:102477 (2021). PubMed: 33937724
- Inoue Y et al. The stimulator of interferon genes (STING) pathway is upregulated in striatal astrocytes of patients with multiple system atrophy. Neurosci Lett 757:135972 (2021). PubMed: 34033888
- Li C et al. RNF111-facilitated neddylation potentiates cGAS-mediated antiviral innate immune response. PLoS Pathog 17:e1009401 (2021). PubMed: 33720974
- Chu L et al. Perillaldehyde Inhibition of cGAS Reduces dsDNA-Induced Interferon Response. Front Immunol 12:655637 (2021). PubMed: 33968056