ZAP 70, phosphorylated (Tyr292) (TCR Zeta Associated Protein kinase 70kD)

Zeta-associated protein (ZAP-70), 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 tyrosine 315 and 319 enhances ZAP-70 function in mediating lymphocyte signaling, while tyrosine 292 terminates transient activation and attentuates lymphocyte signaling. ZAP-70 tyrosine 292 is the negative regulatory site that modulates the duration of activated TCR at the cell surface and functions as a docking site for Cbl\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

Z0300-20

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 Nottini, 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. 3. 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. Ottoson, N.C., et al. (2001) Cutting edge: T cell migration regulated by CXCR4 chemokine receptor signaling to ZAP-70 tyrosine kinase. J. Immunol. 167(4):1857-1861. 4. Lupher, M.L., et al. (1997) The Cbl phosphotyrosine-binding domain selects a D(N/D)XpY motif and binds to the Tyr292 negative regulatory phosphorylation site of ZAP-70. J. Biol. Chem. 272(52):33140-33144. 5. 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. 6. Zhao, Q. and A. Weiss (1996) Enhancement of lymphocyte responsiveness by a gain-of-function mutation of ZAP-70. Mol. Cell. Biol. 16(12):6765-6774.

Additional Info

Recognizes human ZAP-70 when phosphorylated at Tyr292. Species sequence homology: Mouse ZAP-70 (100%).