Product Pathways - Tyrosine Kinase / Adaptors
Phospho-Tie2 (Ser1119) Antibody #4226
PhosphoSitePlus® protein, site, and accession data: TIE2
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W | H | Transfected Only | 160 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 4226:
- Western Blotting
Specificity / Sensitivity
Phospho-Tie2 (Ser1119) Antibody detects transfected levels of Tie2 protein only when phosphorylated at serine 1119.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser1119 of human Tie2. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from Sf9 cells overexpressing GST-human Tie2 kinase domain fusion proteins, wild-type (lane 1) S1119D mutant (lane 2), S1119A mutant (lane 3) or kinase-dead (lane 4) , using Phospho-Tie2 (Ser1119) Antibody (upper) or GST antibody (lower). The wild-type Tie2 kinase domain is constitutively phosphorylated when overexpressed in Sf9 cells. The putative 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).
Phosphorylation of Ser1119 at the carboxy-terminus of Tie2 results in an increase of Tie2 kinase activity (personal communication with Dr. Chris Kontos, Duke University).
- Ward, N.L. and Dumont, D.J. (2002) Semin. Cell Dev. Biol. 13, 19-27.
- Jones, N. and Dumont, D.J. (2000) Cancer Metastasis Rev. 19, 13-17.
- Partanen, J. and Dumont, D.J. (1999) Curr. Top. Microbiol. Immunol. 237, 159-172.
- Ellis, L. M. et al. (2002) Oncology 16, 31-35.
- Koh, G. Y. et al. (2002) Exp. Mol. Med. 34, 1-11.
- Jones, N. et al. (1999) J. Biol. Chem. 274, 30896-30905.
- Jones, N. et al. (2003) Mol. Cell. Biol. 23, 2658-2668.
- Murray, B. W. et al. (2001) Biochem. 40, 10243-10253.
Application References
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For Research Use Only. Not For Use In Diagnostic Procedures.