Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb #2717

Applications Reactivity Sensitivity MW (kDa) Isotype
W H M Endogenous 70, 72 kDa Rabbit IgG

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb detects endogenous levels of Zap-70 only when phosphorylated at tyrosine 319. It cross-reacts with endogenous levels of Syk when phosphorylated at tyrosine 352.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Tyr319 of human Zap-70.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells treated with hydrogen peroxide (2mM for 2 minutes) or with lambda phosphatase and extracts from Ramos cells treated with anti-human IgM (12 micrograms/ml for 2 minutes) using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb.

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 its 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 (CLL) (7,8).

  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.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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