Optimization of FRET Assay with Varioskan® Spectral Scanning Microtiter Plate Fluorometer

Abstract

This paper presents the principle of optimization for fluorometric assays. Optimization is based on the measurement of fluorescence spectra with Thermo Scientific Varioskan® microtitre plate spectral scanning fluorometer. The optimal assay settings for the excitation and emission wavelengths are determined from this spectral data. The optimization procedure is shown with FRET technology based QuantiCleave™ protease assay from Pierce Biotechnology Inc, Rockford, IL (product number 23267) and ApoTarget Caspase-3 assay (product number KHZ0012) from BioSource International Inc, Camarillo, CA.