Isothermal titration calorimetry (ITC) is the method of choice for thermodynamic binding profiling. As a label-free in-solution technique, ITC experiments do not require modified or immobilized proteins or ligands and hence can be performed under close-to-native environmental conditions. It directly follows the released heat of the molecular interaction and does not need competing reporter molecules. However, advanced experimental ITC setups are frequently necessary to derive thermodynamic parameters for medicinal chemistry applications. Beyond 1:1 protein-ligand binding, complex systems include displacement titrations to determine thermodynamics for ligands with affinities either too high or too low to be derived from direct titrations, the characterization of protonation changes upon binding, or the elucidation of heat capacities. We present hands-on experiments for graduate medicinal chemists and students familiar with standard ITC methods, reliably highlighting these issues and their respective solutions. The setup can be either a biochemical trypsin-ligand system or a more cost-efficient EDTA-based variant. This demonstration of best practice protocols comes together with the freely accessible ITC web calculator "ITCcalc" (https://itccalc.uni-mainz.de), guiding through the underlying evaluation of the obtained experimental results.