Isothermal Titration Calorimetry (ITC) is a general technique to characterize binding thermodynamics between two ligands. A single ITC experiment permits the determination of the binding constant, the changes in enthalpy (DH), entropy (DS) and Gibbs free energy (DG). Advantage of ITC over alternative techniques include that almost any type of ligand can be studied and that both ligands are in solution. Furthermore, there are little restrictions as to the choice of the buffer system and the temperature of analysis. In this chapter we provide practical guidelines for the generation of high-quality ITC data giving particular emphasis to trouble shooting. We have used this technique previously to study the binding of hydrocarbons to different macromolecules and information is given which need to be considered when analyzing this class of compound.