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Kinase assay principle. The substrates used in HTRF kinase assays are tagged by fusion proteins, peptides, or biotin. An antibody to these proteins or peptides, or streptavidin (for biotin in this case) is labeled with acceptor. An antibody against phosphorylated substrate is labeled by donor cryptate. In the enzymatic reaction, once substrate is phosphorylated, donor and acceptor can be brought into proximity by affinity interactions of antibodies/substrate or streptavidin/biotin.
