In vitro translation (IVT), or cell-free expression, offers a unique and powerful research tool to screen for translational inhibitors that regulate both cellular and viral protein expression. Here we discuss a simple, quick method to identify inhibitors of both cap-dependent and cap-independent protein translation, using the Thermo Scientific1-Step Human Coupled IVT Kit. Rapid assay readout (6090 min), amenability for miniaturization, and insensitivity to compound toxicity make in vitro translation an attractive alternative to cell-based screens for high-throughput screening (HTS) of novel inhibitors of proteinsynthesis