N-Acetylpuromycin is widely used in biochemical research as a tool to study protein synthesis and ribosome function. It acts as a puromycin analog, inhibiting translation by incorporating into nascent polypeptide chains, causing premature termination. This property makes it valuable for investigating the mechanisms of protein synthesis, ribosome activity, and the effects of translation inhibitors. Additionally, it is employed in assays to monitor cell-free translation systems and to study the fidelity and efficiency of ribosomes. Its ability to halt protein synthesis also aids in identifying newly synthesized proteins and understanding cellular responses to translational stress.