Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-Tie2 (Tyr992) Antibody #4221

Applications Reactivity Sensitivity MW (kDa) Source
W H (M) Transfected Only 160 Rabbit

Applications Key:  W=Western Blotting
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-Tie2 (Tyr992) Antibody detects transfected levels of Tie2 protein only when phosphorylated at tyrosine 992.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr992 of human Tie2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, transfected with wild-type Tie2 (lane 1) or mock-transfected (lane 2), using Phospho-Tie2 (Tyr992) Antibody. Tie2 is constitutively active when transfected into 293 cells.

Western Blotting

Western Blotting

Western blot analysis of extracts from Sf9 cells over expressing GST-human Tie2 kinase domain fusion proteins, wild-type (lane 1) or kinase-dead (lane 2), using Phospho-Tie2 (Tyr992) Antibody (upper) or Tie2 antibody (lower). The wild-type Tie2 kinase domain is constitutively phosphorylated when overexpressed in Sf9 cells. The molecular weight of GST-Tie2 fusion is approximately 65 kDa.

Background

Tie2/Tek is a receptor tyrosine kinase (RTK) expressed almost exclusively on endothelial cells. It is critical for vasculogenesis and could be important for maintaining endothelial cell survival and integrity in adult blood vessels as well as tumor angiogenesis (1-3). A family of ligands known as the angiopoietins binds to Tie2. Interestingly, these ligands appear to have opposing actions; Angiopoietin-1 (Ang1) and Angiopoietin-4 (Ang4) stimulate tyrosine phosphorylation of Tie2; Angiopoietin-2 (Ang2) and Angiopoietin-3 (Ang3) can inhibit this phosphorylation (4,5). Downstream signaling components, including Grb2, Grb7, Grb14, SHP-2, the p85 subunit of phosphatidylinositol 3-kinase, and p56/Dok-2 interact with Tie2 in a phosphotyrosine-dependent manner through their SH2 or PTB domains (6,7). Tyr992 is located on the putative activation loop of Tie2 and is a major autophosphorylation site (8).

  1. Ward, N.L. and Dumont, D.J. (2002) Semin. Cell Dev. Biol. 13, 19-27.
  2. Jones, N. and Dumont, D.J. (2000) Cancer Metastasis Rev. 19, 13-17.
  3. Partanen, J. and Dumont, D.J. (1999) Curr. Top. Microbiol. Immunol. 237, 159-172.
  4. Ellis, L. M. et al. (2002) Oncology 16, 31-35.
  5. Koh, G. Y. et al. (2002) Exp. Mol. Med. 34, 1-11.
  6. Jones, N. et al. (1999) J. Biol. Chem. 274, 30896-30905.
  7. Jones, N. et al. (2003) Mol. Cell. Biol. 23, 2658-2668.
  8. Murray, B. W. et al. (2001) Biochem. 40, 10243-10253.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

Products