The same example data set from pytc was used to analyse the effect of the dilution experiments on the parameter estimation. The four approaches, described in section 3.2.3, were applied: As first approach (1) the titration was analysed with dilution correction, included according to equation 10 but without referring to any external blank titration. Including dilution using another experiment was realised as follows: (2) An external blank titration was used to estimate the two dilution parameters m δ and n δ in equation 10, which were included and kept constants while lgK 11 , ∆H and f x were obtained. The third parameter estimation (3) was performed using equation 8 after the blank experiment was subtracted from the complexation experiment. In the last experiment (4) the dilution and the complexation experiment were combined as MetaModel. Therefore m δ and n δ were estimated using the dilution and the titration experiment globally, while lgK 11 , ∆H and f x were estimated locally, using only the data from the titration experiment. The corresponding isotherm and blank experiment are shown in Fig. , the estimated parameter for hepes-01 are listed in Table . The heat observed from the dilution experiment is very small, compared to the heat from binding experiment. Fig. contains the three isotherms and dilution experiments for the hepes-01, imid-01 and tris-01 data sets. See Tables -S5 for all best fit values as well as the confidence intervals of the parameters lgK 11 and ∆H AB . Monte Carlo simulation with 20000 steps and σ MC = SE y were performed, the corresponding boxplots for lgK 11 and ∆H AB in case