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  1. Link

    naktbk1-antibody-ep611y-ab40676.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases Other Kinases
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-NAK/TBK1 antibody [EP611Y] (ab40676)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (86)

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Western blot - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
  • Western blot - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
  • Immunocytochemistry/ Immunofluorescence - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
  • Western blot - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
  • Immunocytochemistry/ Immunofluorescence - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
  • 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

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Overview

  • 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.

  • 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
  • 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
    For more information see here.

    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

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP611Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Neuroscience
    • Diseases

Associated products

  • Alternative Versions

    • Anti-NAK/TBK1 antibody [EP611Y] - BSA and Azide free (ab186470)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa membrane extract lysate (ab29547)
  • Recombinant Protein

    • Recombinant human NAK/TBK1 protein (ab85276)

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
ICC/IF
1/100.

For unpurified use at 1/250 - 1/500.

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
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

  • 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.
  • Target information above from: UniProt accession Q9UHD2 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • 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 [EP611Y] (ab40676)
      Western blot - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)

      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.

    • Western blot - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
      Western blot - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)

      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)
      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. 

    • Immunocytochemistry/ Immunofluorescence - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
      Immunocytochemistry/ Immunofluorescence - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
      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.
    • Western blot - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
      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
      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 - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
      Immunocytochemistry/ Immunofluorescence - Anti-NAK/TBK1 antibody [EP611Y] (ab40676)

      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.

    • Anti-NAK/TBK1 antibody [EP611Y] (ab40676)
      Anti-NAK/TBK1 antibody [EP611Y] (ab40676)

    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.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (86)

    Publishing research using ab40676? Please let us know so that we can cite the reference in this datasheet.

    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
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-NAK/TBK1 antibody [EP611Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Microglia- CHME3)
    Gel Running Conditions
    Reduced Denaturing (4-16%)
    Loading amount
    15 µg
    Treatment
    250 nM Etoposide for 24hrs
    Specification
    Microglia- CHME3
    Blocking step
    Intercept as blocking agent for 45 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 16 2022

    Western blot abreview for Anti-NAK/TBK1 antibody [EP611Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (MEFs)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    20 µg
    Treatment
    IFN gamma
    Specification
    MEFs
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Prashant Rai

    Verified customer

    Submitted Jul 17 2018

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