|Application ||WB, FC|
|Calculated MW||69872 Da|
|Other Names||Tyrosine-protein kinase ZAP-70, 70 kDa zeta-chain associated protein, Syk-related tyrosine kinase, ZAP70, SRK|
|Target/Specificity||A phospho-specific peptide corresponding to residues surrounding Tyrosine 292 of human ZAP70 was used as an immunogen. This antibody only detects ZAP70 phosphorylated on Tyrosine 292.|
|Format||50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA.|
|Storage||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.|
|Precautions||ZAP70 Antibody Phospho (pY292) is for research use only and not for use in diagnostic or therapeutic procedures.|
|Function||Tyrosine kinase that plays an essential role in regulation of the adaptive immune response. Regulates motility, adhesion and cytokine expression of mature T-cells, as well as thymocyte development. Contributes also to the development and activation of primary B-lymphocytes. When antigen presenting cells (APC) activate T-cell receptor (TCR), a serie of phosphorylations lead to the recruitment of ZAP70 to the doubly phosphorylated TCR component CD247/CD3Z through ITAM motif at the plasma membrane. This recruitment serves to localization to the stimulated TCR and to relieve its autoinhibited conformation. Release of ZAP70 active conformation is further stabilized by phosphorylation mediated by LCK. Subsequently, ZAP70 phosphorylates at least 2 essential adapter proteins: LAT and LCP2. In turn, a large number of signaling molecules are recruited and ultimately lead to lymphokine production, T-cell proliferation and differentiation. Furthermore, ZAP70 controls cytoskeleton modifications, adhesion and mobility of T-lymphocytes, thus ensuring correct delivery of effectors to the APC. ZAP70 is also required for TCR-CD247/CD3Z internalization and degradation through interaction with the E3 ubiquitin-protein ligase CBL and adapter proteins SLA and SLA2. Thus, ZAP70 regulates both T-cell activation switch on and switch off by modulating TCR expression at the T-cell surface. During thymocyte development, ZAP70 promotes survival and cell-cycle progression of developing thymocytes before positive selection (when cells are still CD4/CD8 double negative). Additionally, ZAP70-dependent signaling pathway may also contribute to primary B-cells formation and activation through B-cell receptor (BCR).|
|Cellular Location||Cytoplasm. Cell membrane; Peripheral membrane protein. Note=In quiescent T-lymphocytes, it is cytoplasmic. Upon TCR activation, it is recruited at the plasma membrane by interacting with CD247/CD3Z. Colocalizes together with RHOH in the immunological synapse. RHOH is required for its proper localization to the cell membrane and cytoskeleton fractions in the thymocytes (By similarity).|
|Tissue Location||Expressed in T- and natural killer cells. Also present in early thymocytes and pro/pre B-cells|
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Provided below are standard protocols that you may find useful for product applications.
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).
1. Chu, D.H., et al. Immunol. Rev. 165: 167
2. Isakov, N., et al. J Exp Med. 181: 375
3. Di Bartolo, V., et al. J Biol Chem. 274: 6285
4. G Kong, et al. Mol. Cell. Biol. 16(9):5026-5035, 1996
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