WideScreen RTK Data The WideScreen™ RTK Assays can be applied to study cell signaling as well as the effect of inhibitors on a wide range of human RTKs. The RTK Total Assays enable the quantitation of RTKs for expression profiling, confirming knock-down experiments, and for the normalization of protein levels during inhibitor profiling. The RTK pTyr Assays can be used in secondary screens, to determine the off-target effects of compounds, and have general applications in the analysis of RTK phosphorylation. WideScreen™ RTK Total Assay Standard Curves The RTK Total Standards, Mix, 7-Plex is used to create standard curves to determine the amount of total RTK levels. Recombinant standards are provided for each analyte. Typical standard curves developed with the WideScreen RTK Total Assay Complete Kit are shown.
RTK Expression in Different Cell Lines 
HUVEC and HT-29 cell lysates (10mg total protein) were analyzed with the WideScreen RTK Total Assay Complete Kit to determine expression levels and concentrations of EGFR, IGF-1R, HGFR, PDGFRß, HER-2, VEGFR2, and Tie-2
Inhibitor Profiling 
HT-29 cells were cultured in 96-well plates. Starved cells were incubated with increasing amounts of the EGFR/HER-2 inhibitor PD168393 (Calbiochem Cat. No. 513033) for 1 h and stimulated with a cocktail of growth factors for 10 min. Levels of phosphorylated RTK in cell lysates were tested with the WideScreen RTK pTyr Assay Complete Kit. Levels of phosphorylated protein are shown as percent of positive control (vehicle-treated HT-29 cells). The data show that PD168393 is specific for EGFR/HER-2 and does not inhibit the phosphorylation of IGF-1R or HGFR at sub-micromolar concentrations. HT-29 cells do not express significant amounts of VEGFR2, Tie-2, or PDGFRß. | 
