A Time-resolved Fluorescence Resonance Energy Transfer Assay for Estrogen Receptor Ligands
Various studies have investigated methods to assay the binding affinities of ligands for the estrogen receptor (ER). Although radiometric methods remain the standard, InVitrogen (PanVera) has marketed a fluorescence polarization assay. Still, there is a need for a better, more sensitive, high throughput method. Time-resolved fluorescence resonance energy transfer (TR-FRET) can be utilized in an assay to produce very sensitive readings that can be obtained in a high-throughput manner without the use of radioactivity, and without the need to separate bound ligand from free. The studies reported here will show optimization and validation of the assay by comparing Relative Binding Affinity (RBA) values of some commonly used estrogen ligands obtained by various methods with the current published values. Thus, it will be investigated whether this novel method has the potential to produce reliable data and a means of high-throughput screening, in addition to eliminating hazardous waste.
University of Illinois at Urbana-Champaign
Dr. John A Katzenellenbogen
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