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 (R) (Hm) (Mk) (C) (B) (Dg) (Pg) (Hr) Endogenous 70, 72 kDa Rabbit IgG

Applications Key:  W=Western Blotting
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

Specificity / Sensitivity

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

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide 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 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).

  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.

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

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