Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Phospho-Zap-70 (Tyr493)/Syk (Tyr526) Antibody #2704

Applications Reactivity Sensitivity MW (kDa) Source
W IP F H (M) (R) Endogenous 70 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat
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 (Tyr493)/Syk (Tyr526) Antibody detects endogenous levels of Zap-70 only when phosphorylated at Tyr493. This antibody also cross-reacts with endogenous levels of Syk when phosphorylated at Tyr526.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr493 of human Zap-70. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Ramos cells untreated or treated with 10 µg/ml IgM for 2 minutes with or without calf intestinal phosphatase (CIP), using Phospho-Zap-70 (Tyr493)/Syk (Tyr526) Antibody (upper) or Syk Antibody #12358 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells starved for 16 hours and treated with 2 mM H2O2 for 3 minutes, using Phospho-Zap-70 (Tyr493)/Syk (Tyr526) Antibody (upper) or control Zap-70 Antibody #2702 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or CD3/CD28 treated (green), using Phospho-Zap-70 (Tyr493)/Syk (Tyr526) Antibody compared to a nonspecific negative control antibody (red).


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 Tyr493 within the activation loop results in enzymatic activation of Zap-70 (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. Watts, J.D. et al. (1994) J. Biol. Chem. 269, 29520-29529.

Application References

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

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