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Detection of tyrosine kinase activity in AlphaScreen mode

Franka Ganske, Marjan Orban BMG LABTECH 12/2014

Tyrosine kinases are important regulators of cellular processes and involved in diseases such as cancer and cardiovascular diseases. Therefore, molecules that modulate kinase functions are expected to be promising new drugs. In this application note we will describe the performance of a tyrosine kinase assay using the AlphaScreen® method.

 

AlphaScreen® tyrosine kinase assays performed on the PHERAstar® FS result in consistent values for replicates. A Z' value of 0.93 was calculated, which represents an excellent assay performance. Z' values between 0.5 and 1 indicate a highly robust screening assay and reflect high quality of the instrumentation. Special filters that are optimized for AlphaScreen® enabled a FLUOstar® Omega and a POLARstar® Omega to obtain sensitive and consistent data in 384-well format. The Z'-value for the Omega readers was > 0.8. The limit of detection for PHERAstar®, CLARIOstar® and Omega readers was egal to or greater than 100 amol biot-LCK-P per well.

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