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Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody #2701

Applications	Reactivity	Sensitivity	MW (kDa)	Source
W IP IF-IC F	H (M) (R) (Hm) (Mk) (C) (B) (Dg) (Pg) (Hr)	Endogenous	70 Zap-70, 72 Syk	Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  B=Bovine  Dg=Dog  Pg=Pig  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

2701:

Flow, Immunofluorescence, Immunoprecipitation, Western Blotting

Specificity / Sensitivity

Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody detects endogenous levels of Zap-70 only when phosphorylated at Tyr319. It cross-reacts with endogenous levels of Syk when phosphorylated at Tyr352.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr319 of human Zap-70. Antibodies are purified by protein A and peptide affinity chromatography.

Background

The Syk family protein tyrosine kinase Zap-70 is expressed in T and NK cells and plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement (1). Following TCR engagement, Zap-70 is rapidly phosphorylated on several tyrosine residues through autophosphorylation and transphosphorylation by the Src family tyrosine kinase Lck (2-6). Tyrosine phosphorylation correlates with increased Zap-70 kinase activity and downstream signaling events. Expression of Zap-70 is correlated with disease progression and survival in patients with chronic lymphocytic leukemia (7,8).

Phosphorylation of Tyr319 is required for the assembly of a Zap-70-containing signaling complex that leads to the activation of the PLC-gamma1-dependent and Ras-dependent signaling cascades in antigen-stimulated T cells (5,6). The orthologous Tyr352 residue in Syk is also involved in the association with PLC-gamma1 (9).

1. Chu, D.H. et al. (1998) Immunol. Rev. 165, 167-180.
2. Iwashima, M. et al. (1994) Science 263, 1136-1139.
3. Neumeister, E.N. et al. (1995) Mol. Cell Biol. 15, 3171-3178.
4. Chan, A.C. et al. (1995) EMBO J. 14, 2499-2508.
5. Williams, B.L. et al. (1999) EMBO J. 18, 1832-1844.
6. Di Bartolo, V. et al. (1999) J. Biol. Chem. 274, 6285-6294.
7. Wiestner, A. et al. (2003) Blood 101, 4944-4951.
8. Crespo, M. et al. (2003) N. Engl. J. Med. 348, 1764-1775.
9. Law, C. L. et al. (1996) Mol. Cell. Biol. 16, 1305-1315.

Application References

• Guntermann, C. and Alexander, D.R. (2002) CTLA-4 suppresses proximal TCR signaling in resting human CD4(+) T cells by inhibiting ZAP-70 Tyr(319) phosphorylation: A potential role for tyrosine phosphatases. J. Immunol. 168, 4420-4429. Applications: IP Western Blotting
• Houtman, J. C. et al. (2005) Early phosphorylation kinetics of proteins involved in proximal TCR-mediated signaling pathways. J Immunol. 175 (4), 2449-2458. Applications: Western Blotting
• Schade, A.E. and Levine, A.D. (2002) Lipid Raft Heterogeneity in Human Peripheral Blood T Lymphoblasts: A Mechanism for Regulating the Initiation of TCR Signal Transduction. J. Immunol. 168, 2233-2239. Applications: Western Blotting

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Companion Products

• 2704 Phospho-Zap-70 (Tyr493)/Syk (Tyr526) Antibody
• 2705 Zap-70 (99F2) Rabbit mAb
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 7074 Anti-rabbit IgG, HRP-linked Antibody

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For Research Use Only. Not For Use In Diagnostic Procedures.