While US has been recommended as "one of the most accurate techniques for free-energy calculations [...] only limited by its elevated computational cost", 58 systematic validations are currently lacking. Moreover, the few studies aimed at assessing this validity for specific systems suggest a dependency of the calculated free energies on simulation setup. Despite this controversy, US along a one-dimensional distance-based coordinate has remained a widely accepted method of choice for computing binding affinities, presumably because of its simplicity. To understand if such optimism is justified, and in particular, to assess if the common US workflows are generally valid, we have systematically assessed the accuracy of the US sampling methods for protein-protein and protein-ligand systems.