Guide to Homogeneous Time Resolved Fluorescence

This guide covers the principles and applications of Homogeneous Time Resolved Fluorescence (HTRF), a technique that integrates fluorescence resonance energy transfer with time resolution for sensitive assay development. HTRF employs donor and acceptor fluorophores, facilitating the monitoring of interactions in a homogeneous environment without the need for separation. The document details various detection methods such as ratiometric measurement and outlines the features of HTRF including a no-wash format, low background noise, and application flexibility. It presents a thorough overview of assay formats applicable in both biochemical and cellular contexts, and discusses key components such as the HTRF toolbox reagents and Tag-lite technology. Furthermore, the guide includes a case study on ELISA conversion for glucagon-specific detection. Additionally, it highlights numerous applications in drug discovery, including investigations into GPCRs, kinases, and various biomarkers, while addressing the importance of assay miniaturization and the utility of HTRF in high-throughput screening.