ABSTRACT
Fluorescence has long been recognized to be one of the premiere spectroscopic techniques in sensitivity and breadth of application for probing protein structure for both endogenous and introduced fluorescent molecules (1, 2). Its utility as a biochemical probe in screening has been limited, however, by the very sensitivity that makes it such a powerful technique due to its vulnerability to interfering substances at low concentrations. This is especially true for high throughput screening where the variety of sample types always includes molecules that are fluorescent in their own right (3). Background fluorescence and spurious acceptor molecules which serve to “quench” the emission spectrum readout have combined to make the use of fluorescence detection less than optimal as a screening approach. These problems are more pronounced for natural product samples than for synthetic samples.
