Basic knowledge of equilibrium conditions and the association behavior of any dynamic chemical system is important if one is to evaluate and understand that system. Binding constants for molecular associations can be determined by a variety of different approaches, each with its own advantages and disadvantages. This review examines various chromatographic and electrophoretic methods that have been developed to study dye-protein interactions. An overview of each technique is presented, along with a discussion of its strengths, weaknesses, and potential applications. Examples are provided that illustrate the use of these methods in determining the overall extent of dye-protein binding.