Bodor, Dani L.Rodríguez, Mariluz GómezMoreno, NunoJansen, Lars E.T.2016-05-242016-06-012012-06-01Bodor, D. L., Rodríguez, M. G., Moreno, N. and Jansen, L. E. 2012. Analysis of Protein Turnover by Quantitative SNAP-Based Pulse-Chase Imaging. Current Protocols in Cell Biology. 55:8.8:8.8.1–8.8.34.http://hdl.handle.net/10400.7/614Assessment of protein dynamics in living cells is crucial for understanding their biological properties and functions. The SNAP-tag, a self labeling suicide enzyme, presents a tool with unique features that can be adopted for determining protein dynamics in living cells. Here we present detailed protocols for the use of SNAP in fluorescent pulse-chase and quench-chase-pulse experiments. These time-slicing methods provide powerful tools to assay and quantify the fate and turnover rate of proteins of different ages. We cover advantages and pitfalls of SNAP-tagging in fixed- and live-cell studies and evaluate the recently developed fast-acting SNAPf variant. In addition, to facilitate the analysis of protein turnover datasets, we present an automated algorithm for spot recognition and quantification.engAnimalsCell LineCell SurvivalCentromereFluorescent DyesHumansMicroscopy, FluorescenceProteinsRecombinant Fusion Proteinsjournal article10.1002/0471143030.cb0808s55