FRET-Based Measurement of Apoptotic Caspase Activities by High-Throughput Screening Flow Cytometry

Unwanted and excessive apoptosis contributes to various degenerative diseases, and apoptosis-inducing drugs are a mainstay of anticancer 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 noninvasive, 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.
Source: Springer protocols feed by Pharmacology/Toxicology - Category: Drugs & Pharmacology Source Type: news