This dataset includes isothermal titration calorimetry measurements of human thymidylate synthase (hTS) binding its product deoxythymidine monophosphate (TMP) and an N-terminal trunction of hTS binding its substrate deoxyuridine monophosphate (dUMP). These measurements include heats produced at various enzyme and ligand concentrations. Analysis of these data provide binding free energies, enthalpies, and entropies for these binding events. We find that in contrast to hTS binding its substrate dUMP, which we have previously shown to have positive cooperativity, hTS binding TMP and the N-terminally truncated hTS binding dUMP are non-cooperative. Details of the data collection and analysis can be found in the associated publication.