Real-time monitoring of clathrin assembly kinetics in a reconstituted system

Abstract

Clathrin-coated pits (ccp) are important structures that cells use for internalizing materials and regulating plasma membrane homeostasis. We had previously described an assay of reconstituting ccp assembly on sheets of basal plasma membranes. Here, we describe a workflow to adapt this system for monitoring the assembly of ccps over time using total internal reflection fluorescence (TIRF) microscopy.