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

Recombinant Anti-TAK1 antibody [EPR5984] (ab109526)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (34)

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Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)
  • Immunocytochemistry/ Immunofluorescence - Anti-TAK1 antibody [EPR5984] (ab109526)
  • Flow Cytometry - Anti-TAK1 antibody [EPR5984] (ab109526)
  • Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)
  • Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAK1 antibody [EPR5984] (ab109526)
  • Anti-TAK1 antibody [EPR5984] (ab109526)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5984] to TAK1
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-TAK1 antibody [EPR5984]
    See all TAK1 primary antibodies
  • Description

    Rabbit monoclonal [EPR5984] to TAK1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293T, K562, HeLa and A431 cell lysates. IHC-P: Human brain tissue. ICC/IF: Wild-type HAP1 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
    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.

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. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR5984
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Serine/threonine kinases
    • MAPK pathway
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Immunology
    • Innate Immunity
    • TLR Signaling
    • Cancer
    • Cell Death
    • Necroptosis

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-TAK1 antibody [EPR5984] (ab199191)
    • Alexa Fluor® 647 Anti-TAK1 antibody [EPR5984] (ab199473)
    • Anti-TAK1 antibody [EPR5984] - BSA and Azide free (ab222394)
  • 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)
  • KO cell lines

    • Human MAP3K7 (TAK1) knockout HEK-293T cell line (ab266555)
  • KO cell lysates

    • Human MAP3K7 (TAK1) knockout HEK-293T cell lysate (ab256984)
  • KO cell pellets

    • Human MAP3K7 (TAK1) knockout HEK-293T cell pellet (ab278834)
  • Recombinant Protein

    • Recombinant human TAK1 protein (ab132950)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109526 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 (2)
1/1000 - 1/10000. Detects a band of approximately 75 kDa (predicted molecular weight: 67 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol. (Heat to 98°C, allow to cool for 10-20 minutes)
ICC/IF
1/1000.
Flow Cyt
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB
1/1000 - 1/10000. Detects a band of approximately 75 kDa (predicted molecular weight: 67 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol. (Heat to 98°C, allow to cool for 10-20 minutes)
ICC/IF
1/1000.
Flow Cyt
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Component of a protein kinase signal transduction cascade. Mediator of TRAF6 and TGF-beta signal transduction. Activates IKBKB and MAPK8 in response to TRAF6 signaling. Stimulates NF-kappa-B activation and the p38 MAPK pathway. In osmotic stress signaling, plays a major role in the activation of MAPK8/JNK, but not that of NF-kappa-B.
    • Sequence similarities

      Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications

      Association with TAB1/MAP3K7IP1 promotes autophosphorylation and subsequent activation. Association with TAB2/MAP3K7IP2, itself associated with free unanchored Lys-63 polyubiquitin chain, promotes autophosphorylation and subsequent activation of MAP3K7. Dephosphorylation at Thr-187 by PP2A and PPP6C leads to inactivation.
      Ubiquitinated, leading to proteasomal degradation (By similarity). Requires 'Lys-63'-linked polyubiquitination for autophosphorylation and subsequent activation. 'Lys-63'-linked ubiquitination does not lead to proteasomal degradation. Deubiquitinated by CYLD, a protease that selectively cleaves 'Lys-63'-linked ubiquitin chains. Deubiquitinated by Y.enterocolitica YopP.
    • Target information above from: UniProt accession O43318 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 6885 Human
      • Entrez Gene: 26409 Mouse
      • Entrez Gene: 313121 Rat
      • Omim: 602614 Human
      • SwissProt: O43318 Human
      • SwissProt: Q62073 Mouse
      • SwissProt: P0C8E4 Rat
      • Unigene: 594838 Human
      • Unigene: 722892 Human
      • Unigene: 258589 Mouse
      • Unigene: 24019 Rat
      see all
    • Alternative names

      • M3K7_HUMAN antibody
      • MAP3K 7 antibody
      • Map3k7 antibody
      • MEKK7 antibody
      • Mitogen activated protein kinase kinase kinase 7 antibody
      • Mitogen-activated protein kinase kinase kinase 7 antibody
      • TAK1 antibody
      • TGF beta activated kinase 1 antibody
      • TGF-beta-activated kinase 1 antibody
      • TGF1a antibody
      • Transforming growth factor beta activated kinase 1 antibody
      • Transforming growth factor-beta-activated kinase 1 antibody
      see all

    Images

    • Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)
      Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: TAK1 knockout HAP1 cell lysate (20 µg)
      Lane 3: SHSY5Y cell lysate (20 µg)
      Lane 4: A431 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab109526 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab109526 was shown to specifically react with TAK1 when TAK1 knockout samples were used. Wild-type and TAK1 knockout samples were subjected to SDS-PAGE. ab109526 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-TAK1 antibody [EPR5984] (ab109526)
      Immunocytochemistry/ Immunofluorescence - Anti-TAK1 antibody [EPR5984] (ab109526)

      ab109526 staining TAK1 in wild-type HAP1 cells (top panel) and TAK1 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109526 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Flow Cytometry - Anti-TAK1 antibody [EPR5984] (ab109526)
      Flow Cytometry - Anti-TAK1 antibody [EPR5984] (ab109526)

      Flow Cytometry analysis of A431(human epidermoid carcinoma) cells labeling TAK1 with unpurified ab109526 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

    • Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)
      Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)
      All lanes : Anti-TAK1 antibody [EPR5984] (ab109526) at 1/1000 dilution

      Lane 1 : K562 cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : A431 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution

      Predicted band size: 67 kDa
      Observed band size: 75 kDa why is the actual band size different from the predicted?

    • Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)
      Western blot - Anti-TAK1 antibody [EPR5984] (ab109526)
      All lanes : Anti-TAK1 antibody [EPR5984] (ab109526) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : MAP3K7 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 67 kDa
      Observed band size: 72 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab109526 observed at 72 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab109526 was shown to react with TAK1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266555 (knockout cell lysate ab256984) was used. Wild-type HEK-293T and MAP3K7 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109526 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAK1 antibody [EPR5984] (ab109526)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAK1 antibody [EPR5984] (ab109526)

      ab109526, at a 1/50 dilution, staining TAK1 in Formalin/PFA-fixed paraffin-embedded Human brain tissue, by Immunohistochemistry.

      Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.

    • Anti-TAK1 antibody [EPR5984] (ab109526)
      Anti-TAK1 antibody [EPR5984] (ab109526)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (34)

    Publishing research using ab109526? Please let us know so that we can cite the reference in this datasheet.

    ab109526 has been referenced in 34 publications.

    • Xu M  et al. Functional loss of inactive rhomboid-like protein 2 mitigates obesity by suppressing pro-inflammatory macrophage activation-triggered adipose inflammation. Mol Metab 34:112-123 (2020). PubMed: 32180551
    • Rong X  et al. Enalapril attenuates endoplasmic reticulum stress and mitochondrial injury induced by myocardial infarction via activation of the TAK1/NFAT pathway in mice. Exp Ther Med 19:972-980 (2020). PubMed: 32010259
    • Yang G & Zhao Y Overexpression of miR-146b-5p Ameliorates Neonatal Hypoxic Ischemic Encephalopathy by Inhibiting IRAK1/TRAF6/TAK1/NF-aB Signaling. Yonsei Med J 61:660-669 (2020). PubMed: 32734729
    • Zeng J  et al. Inhibition of TGFß-activated protein kinase 1 ameliorates myocardial ischaemia/reperfusion injury via endoplasmic reticulum stress suppression. J Cell Mol Med 24:6846-6859 (2020). PubMed: 32378287
    • Xu K  et al. Protective Effects of SIRT6 Overexpression against DSS-Induced Colitis in Mice. Cells 9:N/A (2020). PubMed: 32580272
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review

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    1-2 of 2 Abreviews

    Western blot abreview for Anti-TAK1 antibody [EPR5984]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (hepatocytes)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    hepatocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 10 2015

    Western blot abreview for Anti-TAK1 antibody [EPR5984]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    40 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (Hep3B)
    Specification
    Hep3B
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 17 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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