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Isothermal Titration Calorimetry (ITC) is a label-free quantification technique used in studies of a wide variety of biomolecular interactions. It works by directly measuring the heat that is either released or absorbed during a biomolecular binding event. ITC is used to investigate all types of protein interactions, including protein-protein interactions, protein-DNA/RNA interactions, protein-small molecule interactions and enzyme kinetics; it provides a direct route to the complete thermodynamic characterization of protein interactions. A key advantage of using ITC for measuring bimolecular interactions is that it can be used on free molecules in solution.