The measurement of equilibrium binding constants for ligand-macromolecule interactions by monitoring a change in some spectral property of the ligand or the macromolecule is a common method used to study these interactions. This is due to the high sensitivity of the spectroscopic methods and general ease in applying these experimental procedures. In addition, binding can be monitored continuously, thus facilitating kinetic measurements. The main problem with these methods results from the fact that the spectroscopic signal is an indirect measure of binding, since the relationship between the change in the spectroscopic signal and the extent of binding is unknown, a priori. A common recourse is to assume a strict proportionality between the signal change and the fractional saturation of the ligand or macromolecule; however, it is often the case that such a direct proportionality does not hold. In this chapter we have reviewed the use of methods to analyze ligand-macromolecule equilibrium titrations that are monitored by indirect spectroscopic techniques. These methods of analysis yield thermodynamically rigorous, model-independent binding isotherms, hence assumptions concerning the relationship between the signal change and the extent of binding are not required. In fact, these methods can also be used to determine quantitatively the relationship between the signal change and the average degree of binding. In addition, the approaches discussed here are general and not limited to spectroscopic signals and therefore can be used with any intensive physicochemical property that reflects binding.