Fig. 1.

Simulated binding isotherm for a 1:1 system to show the importance of c value. The data were generated using the model proposed by Wiseman et al. [10] using ΔH = −50 kcal/mol. For low c values (<5), the binding isotherm loses the S-shape and thus determination of the thermodynamic parameters is not possible. At high c values (∼1,000), ΔH and stoichiometry (n) can be inferred from the shape of the binding isotherm but the data cannot be fitted to determine the other thermodynamic parameters