A strategy to take advantage of time-resolved luminescence of Ln3+-chelated phosphotyrosine-containing peptides, which facilitate efficient energy transfer to small molecule fluorophores conjugated to the peptides to produce orthogonally-colored biosensors for two different kinases is provided. The method enables multiplexed detection with high signal to noise in a high-throughput-compatible format and a platform that could be applied to other lanthanide metal and fluorophore combinations to achieve even greater multiplexing without the need for phosphospecific antibodies.
KINASE ACTIVITY DETECTION METHODS