Imaging Secretory Granule Budding from the Trans-Golgi Network Using Retention Using Selective Hook (RUSH)

The retention using selective hook (RUSH) system enables us to synchronize and visualize the movement of cargoes along the secretory pathway. A fluorescently tagged cargo of interest is retained in the endoplasmic reticulum and released in a biotin-dependent manner. Here, we report a detailed protocol describing the steps necessary to perform RUSH experiments to study secretory granule biogenesis at the trans-Golgi network (TGN).