This resource from Enzo Life Sciences offers practical advice and best practices for developing successful cell-based assays. Cell-based assays are crucial in drug development programs for evaluating compound effects on cellular targets, assessing cytotoxicity, and monitoring proliferation. The tips emphasize critical considerations such as choosing appropriate cell types, using suitable culture media and supplements, regulating incubator conditions (temperature and CO2), and avoiding contamination. It also highlights the importance of ensuring cells are healthy and viable, and minimizing exposure to toxic agents like trypsin during cell dissociation. Consistency in experimental procedures, including using the same stock of cells and controlling cell confluency, is stressed to minimize variability and ensure reproducibility. The resource helps researchers design assays that faithfully represent the biological system and express necessary factors and signaling intermediates. These guidelines are essential for anyone working with cell cultures, from beginners to experienced practitioners, aiming to improve daily tasks and the overall quality of their research by ensuring robust and reliable assay performance.