Paul Held Ph. D., Xavier Amouretti, Fabienne Chevallier, Alexandre Jean
BioTek Instruments, Winooski , VT, USA. Cisbio, Codolet, France
cAMP and Tumor Necrosis Factor (TNF-?) are important mediators and indicators for a litany of cellular responses. As such, assays of these moieties are common in research and drug discovery, including high throughput screening (HTS). The ability to accurately measure changes in these compounds, using a homogeneous assay technology, saves considerable amounts of time and expense. High-throughput assays require that both the assay technology, as well as the instrumentation necessary to measure the results, be sensitive, reliable and cost effective. Assays based on Cisbio's Homogeneous Time-Resolved Fluorescence (HTRF®) technology use a combination of Time-Resolved Fluorescence (TRF) and Fluorescence Resonance Energy Transfer (FRET) to investigate biomolecular interactions. BioTek's Synergy 2 Multi-Detection Microplate Reader combines a high intensity xenon flash lamp with dichroic mirrors and deep blocking fluorescence filters to provide exquisite sensitivity. The combination of a high performance multi-detection reader and a robust homogeneous assay technology provides for a reliable HTS solution. Here we describe the use of the Synergy 2, in conjunction with Gen5 Data Analysis Software, to quantitate the signal and perform the data reduction for cAMP and TNF assays using HTRF® technology from Cisbio.