FRET-based measurement of apoptotic caspase activities by high-throughput screening flow cytometry
Christian T. Hellwig, Agnieszka H. Ludwig-Galezowska, and Markus Rehm
in Apoptosis Methods in Toxicology, Humana Press/Springer; DOI 10.1007/978-01-4939-3588-8_7
Unwanted and excessive apoptosis contributes to various degenerative diseases, and apoptosis-inducing drugs are a mainstay of anti-cancer treatment regimens. The fields of pharmacology and toxicology consequently have a long history of investigating apoptotic cell death in the context of drug safety and efficacy studies. Canonical apoptotic cell death is crucially dependent on type II cysteinyl aspartate-specific proteases (caspases), and their activation is therefore widely used as a marker for this cell death modality. Here we describe a flow cytometric method for non-invasive, highly sensitive and reproducible FRET-based measurements of caspase activation. Compared to other flow cytometric techniques for apoptosis detection, this approach requires only minimal sample handling steps and provides a highly cost efficient option for large scale drug interaction studies and screens of compound libraries.