ABSTRACT
HTRF chemistry was developed to help overcome some of the obstacles to HTS in general. It is homogeneous, so that separation steps are eliminated. It is fluorescent in nature but has the time-resolved, simultaneous, dual-wavelength detection that enables one to achieve high fluorescent signal with extremely low backgrounds. HTRF is best measured on the Discovery HTRF microplate analyzer because it was designed specifically and optimized for the particular wavelengths of HTRF. However, a number of detection systems are now available for measuring TRF and HTRF applications. The HTRF reagent list is growing; thus, fewer assays require customized conjugation with (Eu)K and XL665. Many of the applications described utilize off-the-shelf reagents that are readily available and require no license fee. As a result, this tool is widely accepted in the literature as one of the few truly homogeneous methods available to meet the HTS demands of drug discovery today.
