Overview

  • Product name

    Anti-ITK/EMT antibody [Y401]
    See all ITK/EMT primary antibodies
  • Description

    Rabbit monoclonal [Y401] to ITK/EMT
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human ITK/EMT aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Jurkat whole cell lysate (ab7899). IHC-P: Human lymph node.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y401
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32039 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 1/1000 - 1/5000. Detects a band of approximately 68 kDa (predicted molecular weight: 72 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt 1/100.

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

  • Application notes
    Is unsuitable for IP.
  • 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

    • Anti-ITK/EMT antibody [Y401] (ab32039) at 1/5000 dilution + Jurkat cell lysate

      Predicted band size: 72 kDa
      Observed band size: 68 kDa
      why is the actual band size different from the predicted?

    • Ab32039 at a 1/250 dilution, staining ITK/EMT in paraffin embedded human lymph node tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Overlay histogram showing Jurkat cells stained with ab32039 (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. The cells were then incubated with the antibody (ab32039, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (1µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    Protocols

    References

    This product has been referenced in:

    • Liu Y  et al. ITK inhibition induced in vitro and in vivo anti-tumor activity through downregulating TCR signaling pathway in malignant T cell lymphoma. Cancer Cell Int 19:32 (2019). Read more (PubMed: 30814910) »
    • Mamand S  et al. Comparison of interleukin-2-inducible kinase (ITK) inhibitors and potential for combination therapies for T-cell lymphoma. Sci Rep 8:14216 (2018). Read more (PubMed: 30242208) »
    See all 5 Publications for this product

    Customer reviews and Q&As

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (T-cells - Jurkat line)
    Permeabilization
    Yes - 0.1% Triton X-100 in PBS, room temp., 15 minutes
    Specification
    T-cells - Jurkat line
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Formaldehyde

    Ethan Strattan

    Verified customer

    Submitted Dec 27 2017

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