ZAP 70, phosphorylated (Tyr315, Tyr319) (TCR Zeta Associated Protein kinase 70kD)

Zeta-associated protein (ZAP-70, TCR zeta associated protein kinase 70kD), a 70kD member of the Syk tyrosine kinase family, plays a central role in lymphocyte activation and development and is implicated in several immune disorders. Upon T-cell antigen receptor (TCR) engagement, ZAP-70 is phosphorylated on tyrosines 292, 315 and 319 in the interdomain B, located between the SH2 and kinase domains. Phosphorylation of both tyrosines 315 (a Vav-binding site) and 319 (a Lck binding site) enhances ZAP-70 function in mediating lymphocyte signaling, while tyrosine 292 terminates the transient activation of ZAP-70 and attentuates lymphocyte signaling. Phosphorylation of tyrosines 315 and 319 plays an important role in mediating the positive and negative selection of T cells in thymus.\n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000 \nOptimal dilutions to be determined by the researcher.\n\nPositive Control:\nJurkat cells treated with H2O2\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

Z0300-22

Size

10 Blots

Applications

WB

Hosts

Rabbit

Reactivities

Hum

Form

Supplied as a liquid in PBS (without Mg2+ and Ca2+), 1mg/ml BSA (IgG, protease free), pH 7.2, 0.05% sodium azide, 50% glycerol.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

References

1. Orchard, J.A., et al. (2004) ZAP-70 expression and prognosis in chronic lymphocytic leukaemia. Lancet 363(9403):105-111. 2. Sakaguchi, N., et al. (2003) Altered thymic T-cell selection due to a mutation of the ZAP-70 gene causes autoimmune arthritis in mice Nature 426(6965):454-460. 3. Bottini, N., et al. (2002) Activation of ZAP-70 through specific dephosphorylation at the inhibitory Tyr-292 by the low molecular weight phosphotyrosine phosphatase (LMPTP). J. Biol. Chem. 277(27):24220-24224. 4. Di Bartolo, V., et al. (2002) Tyrosine 315 determines optimal recruitment of ZAP-70 to the T cell antigen receptor. Eur. J. Immunol. 32(2):568-575. 5. Magnan, A., et al. (2001) T cell development and T cell responses in mice with mutations affecting tyrosines 292 or 315 of the ZAP-70 protein tyrosine kinase. J. Exp. Med. 194(4):491-505. Gong, Q., et al. (2001) Requirement for tyrosine residues 315 and 319 within zeta chain-associated protein 70 for T cell development. J. Exp. Med. 194(4):507-518. 6. Wu, J., et al. (1997) The Vav binding site (Y315) in ZAP-70 is critical for antigen receptormediated signal transduction. J. Exp. Med. 185(10):1877-1882. 7. Kong, G., et al. (1996) Distinct tyrosine phosphorylation sites in ZAP-70 mediate activation and negative regulation of antigen receptor function. Mol. Cell. Biol. 16(9):5026-5035.

Additional Info

Recognizes human ZAP-70 when phosphorylated at Tyr315 and Tyr319. Species sequence homology: Mouse ZAP-70 (100%), Syk (85%).