TIE2 Cellular Phosphorylation Assay Service

TIE2 Cellular Phosphorylation Assay (intracellular kinase activity assay) for compound screening and profiling in intact cells

Target Overview

Target Description

The angiopoietin (Ang)-Tie ligand-receptor system has a key role in regulating vascular integrity and quiescence. Besides its role in angiogenesis it is an important regulator in numerous diseases including inflammation. Ang-1-mediated Tie2 activation is required to maintain the quiescent resting state of the endothelium. In contrast, Ang-2 destabilizes the quiescent endothelium and primes it to respond to exogenous stimuli, thereby facilitating the activities of inflammatory and angiogenic cytokines. Intriguingly, Ang-2 is expressed weakly by the resting endothelium but becomes strongly upregulated following endothelial activation. Moreover, endothelial cells store Ang-2 in Weibel-Palade bodies from where it can be made available quickly following stimulation, suggesting a role of Ang-2 in controlling rapid vascular adaptive processes. This suggests that Ang-2 functions as a built-in switch controlling the transition of the resting quiescent endothelium towards the activated responsive endothelium (Trends Immunol. 2006 27, p552-8).

HGNC Reference




Cell Line


Target Expression


Assay Properties

Assay Description

CHO cells have been stably transfected to overexpress full length human TIE2. Stimulation of these cells with
sodium orthovanadate results in a robust receptor autophosphorylation. Compounds are preincubated before cell stimulation to allow thorough target binding. Stimulation conditions are optimized to determine dose-related inhibition of the phospho-TIE2 signal, which is subsequently quantified by Sandwich-ELISA technique. The assay is validated based on known inhibitors of TIE2 kinase activity (see Fig. 1).


Substrate phosphorylation as a readout of intracellular kinase activity via ELISA

Screening Location

Freiburg, Germany

Further Information

More information can be found on our website Cellular Phosphorylation Assay Services.

Reference compound IC50 for TIE2

Reference compound IC50 for TIE2

Sorafenib is a potent inhibitor of TIE2 phosphorylation with highly reproducible IC50 values. The graph shows a representative result.