Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y402] to ITK/EMT
- Suitable for: WB, IP
- Reacts with: Human
Product nameAnti-ITK/EMT antibody [Y402]
See all ITK/EMT primary antibodies
DescriptionRabbit monoclonal [Y402] to ITK/EMT
Tested applicationsSuitable for: WB, IPmore details
Unsuitable for: Flow Cyt,ICC/IF or IHC
Species reactivityReacts with: Human
Synthetic peptide within Human ITK/EMT (C terminal). The exact sequence is proprietary.
- Jurkat whole cell lysate (ab7899).
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.
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.
Storage instructionsShipped 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 bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.21% BSA
Concentration information loading...
PurityProtein A purified
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.
|WB||1/10000. Detects a band of approximately 72 kDa (predicted molecular weight: 72 kDa).
For unpurified use at 1/1000.
For unpurified use at 1/50.
FunctionPlays a role in T-cell proliferation and differentiation.
Tissue specificityT-cell lines and natural killer cell lines.
Involvement in diseaseDefects 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 similaritiesBelongs 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 localizationCell membrane. Localizes to cell surface receptors in the plasma membrane after stimulation with respective receptors (TCR, CD28, CD2) in T-cells.
- Information by UniProt
- EMT antibody
- Homolog of mouse T cell itk/tsk antibody
- IL 2 inducible T cell kinase antibody
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.
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
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.
ab32507 has been referenced in 2 publications.
- 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). 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). PubMed: 19425169