High-throughput screening (HTS) is a crucial technique for identifying potential hits to fuel drug discovery pipelines. However, this process naturally concentrates nuisance compounds that are not optimizable yet signal positively in a convincing manner. To be able to understand what types of nuisance compounds a particular assay is sensitive to, would be of great utility in being able to prioritize progressable over nonprogressable screening hits. In this study, we present a carefully compiled set of over 100 nuisance compounds that are known to interfere with assay readouts in either phenotypic or target-based screenings. Readily accessible in an assay-ready screening plate, we believe this nuisance compound set will be of great interest to the research community, helping to establish high-quality HTS assays and identify promising, optimizable hits.