Purification of clathrin-coated vesicles from adult rat brain

Abstract

Here, we describe a purification protocol for isolating clathrin-coated vesicles (CCVs) from adult rat brain by using differential centrifugation coupled with Ficoll–sucrose and D2O–sucrose density gradient centrifugation and an additional linear sucrose step gradient at the end to separate CCVs from contaminating membranes present in the crude microsomal fraction.