Overview

  • Product name

    Anti-ITK/EMT antibody [Y402]
    See all ITK/EMT primary antibodies
  • Description

    Rabbit monoclonal [Y402] to ITK/EMT
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human ITK/EMT (C terminal). The exact sequence is proprietary.

  • Positive control

    • Jurkat whole cell lysate (ab7899).
  • General notes

     Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Previously labelled as ITK. 

    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.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.21% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y402
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32507 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 1/10000. Detects a band of approximately 72 kDa (predicted molecular weight: 72 kDa).

For unpurified use at 1/1000.

IP 1/20.

For unpurified use at 1/50.

  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC.
  • Target

    • Function

      Plays a role in T-cell proliferation and differentiation.
    • Tissue specificity

      T-cell lines and natural killer cell lines.
    • Involvement in disease

      Defects in ITK are the cause of lymphoproliferative syndrome EBV-associated autosomal type 1 (LPSA1) [MIM:613011]. LPSA1 is a rare immunodeficiency characterized by extreme susceptibility to infection with Epstein-Barr virus (EBV). Inadequate immune response to EBV can have a fatal outcome. Clinical features include splenomegaly, lymphadenopathy, anemia, thrombocytopenia, pancytopenia, recurrent infections. There is an increased risk for lymphoma.
    • Sequence similarities

      Belongs to the protein kinase superfamily. Tyr protein kinase family. TEC subfamily.
      Contains 1 Btk-type zinc finger.
      Contains 1 PH domain.
      Contains 1 protein kinase domain.
      Contains 1 SH2 domain.
      Contains 1 SH3 domain.
    • Cellular localization

      Cell membrane. Localizes to cell surface receptors in the plasma membrane after stimulation with respective receptors (TCR, CD28, CD2) in T-cells.
    • Information by UniProt
    • Database links

    • Alternative names

      • EMT antibody
      • Homolog of mouse T cell itk/tsk antibody
      • IL 2 inducible T cell kinase antibody
      • IL2 inducible T cell kinase antibody
      • Interleukin 2 inducible T cell kinase antibody
      • Itk antibody
      • ITK_HUMAN antibody
      • Kinase EMT antibody
      • LPFS1 antibody
      • LYK antibody
      • MGC126257 antibody
      • MGC126258 antibody
      • PSCTK 2 antibody
      • PSCTK2 antibody
      • T cell specific kinase antibody
      • T-cell-specific kinase antibody
      • TSK antibody
      • Tyrosine protein kinase antibody
      • Tyrosine protein kinase ITK/TSK antibody
      • Tyrosine protein kinase Lyk antibody
      • Tyrosine-protein kinase ITK/TSK antibody
      • Tyrosine-protein kinase Lyk antibody
      see all

    Images

    • Lane 1 : Anti-ITK/EMT antibody [Y402] (ab32507) at 1/2000 dilution (purified)
      Lane 2 : Anti-ITK/EMT antibody [Y402] (ab32507) at 1/10000 dilution (purified)

      All lanes : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 72 kDa



      Blocking and diluting buffer: 5% NFDM/TBST

    • ab32507 (purified) at 1:20 dilution (1.0μg) immunoprecipitating ITK/EMT in Jurkat whole cell lysate.

      Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate, 10μg
      Lane 2 (+): ab32507 & Jurkat whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab172638 in Jurkat whole cell lysate

      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Overlay histogram showing Jurkat cells stained with unpurified ab32507 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32507, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Anti-ITK/EMT antibody [Y402] (ab32507) at 1/1000 dilution (unpurified) + Jurkat cell lysate

      Predicted band size: 72 kDa
      Observed band size: 72 kDa

    References

    This product has been referenced in:

    • Patel V  et al. Comparison of Acalabrutinib, A Selective Bruton Tyrosine Kinase Inhibitor, with Ibrutinib in Chronic Lymphocytic Leukemia Cells. Clin Cancer Res 23:3734-3743 (2017). Read more (PubMed: 28034907) »
    • Huck K  et al. Girls homozygous for an IL-2-inducible T cell kinase mutation that leads to protein deficiency develop fatal EBV-associated lymphoproliferation. J Clin Invest 119:1350-8 (2009). Read more (PubMed: 19425169) »
    See all 2 Publications for this product

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