Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Phospho-Syk (Tyr323) Antibody #2715

Applications Reactivity Sensitivity MW (kDa) Source
W IP H (M) Endogenous 72 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse
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-Syk (Tyr323) Antibody detects endogenous levels of Syk only when phosphorylated at Tyr323. It does not cross-react with other tyrosine-phosphorylated members of the Syk/Zap-70 tyrosine kinase family.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr323 of human Syk or Tyr317 of mouse Syk. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells expressing recombinant wild-type or mutant Syk proteins, cotransfected with CD8, using Phospho-Syk (Tyr323) Antibody (upper) or Syk Antibody #2712 (lower). (Provided by Dr. Alagarsamy L. Reddi, laboratory of Dr. Hamid Band, Harvard University, Massachusetts.)

Western Blotting

Western Blotting

Western blot analysis of extracts from Ramos cells, untreated or treated with anti-human-IgM (10 µg/ml), using Phospho-Syk (Tyr323) Antibody #2715 (upper) or Syk Antibody #2712 (lower).

Background

Syk is a protein tyrosine kinase that plays an important role in intracellular signal transduction in hematopoietic cells (1-3). Syk interacts with immunoreceptor tyrosine-based activation motifs (ITAMs) located in the cytoplasmic domains of immune receptors (4). It couples the activated immunoreceptors to downstream signaling events that mediate diverse cellular responses, including proliferation, differentiation and phagocytosis (4). There is also evidence of a role for Syk in nonimmune cells, and Syk is a potential tumor suppressor in human breast carcinomas (5). Tyr323 is a negative regulatory phosphorylation site within the SH2-kinase linker region in Syk. Phosphorylation of Tyr323 provides a direct binding site to the TKB domain of Cbl (6,7). Tyrosine 352 of Syk is involved in the association of PLC-γ1 (8). Tyrosines 525 and 526 are located in the activation loop of the Syk kinase domain, and phosphorylation of Tyr525/526 of human Syk (equivalent to the Tyr519/520 of mouse Syk) is essential for Syk function (9).

  1. Cheng, A.M. and Chan, A.C. (1997) Curr. Opin. Immunol. 9, 528-533.
  2. Kurosaki, T. et al. (1997) Curr. Opin. Immunol. 9, 309-318.
  3. Chu, D.H. et al. (1998) Immunol. Rev. 165, 167-180.
  4. Turner, M. et al. (2000) Immunol. Today 21, 148-154.
  5. Coopman, P.J. et al. (2000) Nature 406, 742-747.
  6. Deckert, M. et al. (1998) J. Biol. Chem. 273, 8867-8874.
  7. Rao, N. et al. (2001) EMBO J. 20, 7085-7095.
  8. Law, C.L. et al. (1996) Mol. Cell. Biol. 16, 1305-1315.
  9. Zhang, J. et al. (2000) J. Biol. Chem. 275, 35442-35447.

Application References

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Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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