Fluorescence polarization/anisotropy approaches to study protein-ligand interactions: effects of errors and...

Fluorescence techniques are widely used in the study of protein-ligand interactions because of their inherent sensitivity, and the fact that they can be implemented at true equilibrium conditions. Fluorescence polarization/anisotropy methodologies, in particular, are now extensively utilized in biotechnology and clinical chemistry. In this chapter, we shall discuss both theoretical and practical aspects of polarization/anisotropy methods. We shall also focus attention on considerations of errors and uncertainties in such measurements, and how these uncertainties affect the ultimate estimation of ligand-protein dissociation constants.