ZAP70, a Syk-family protein tyrosine kinase (PTK), plays a critical role in mediating T cell signal
transduction in response to T cell receptor (TCR) activation 1). TCR-mediated activation of the Src-family kinases, Lck and Fyn, results in tyrosine phosphorylation of the TCR zeta and CD3 chains. These domains serve as targets for binding of ZAP70 via its tandem SH2 domains. This binding correlates with activation of ZAP70, a critical event in T cell activation (2). Following TCR engagement, ZAP70 is phosphorylated on several tyrosine residues, presumably by two mechanisms: an autophosphorylation and a trans- phosphorylation by the Src-family tyrosine kinase, Lck (3). Heterologous trans- phosphorylation of Tyrosine 493 by Src- PTKs is required for antigen receptor-mediated activation of both the calcium and Ras pathways. In contrast however, cells expressing mutations at Tyrosine 292 or 492 demonstrate hyperactive T- and B-cell antigen receptor phenotypes. Thus, phosphorylation of ZAP-70 mediates both activation and inactivation of antigen receptor signaling (4).
Applications:
Suitable for use in Flow Cytometry, Western Blot, Immunocytochemistry. Other applications not tested.
Recommended Dilution:
Western Blot: 1:5000-10,000
Immunocytochemistry: 1:250-1:500
Flow cytometry: 1:20
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Manufactured incorporating RabMAb(R) technology under Epitomics US patents, No 5,675,063 and 7,429,487, owned by Abcam.
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