title: Preparation of enriched plant clathrin-coated vesicles by differential and density gradient centrifugation.
abstract: Methods for the subcellular fractionation and enrichment of specific intracellular compartments are essential tools in the analysis of compartment composition and function. In vitro characterization of isolated cell organelles and other endomembrane intermediates, including exploration of the compartment protein ensemble, offers strong clues of in vivo function identity. Here, we describe methodology for the isolation of clathrin-coated vesicles from Arabidopsis thaliana suspension-cultured cells on the basis of differential and density centrifugation.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Dr., Madison, WI, 53706, USA.
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Reynolds
authors.forename: Ben
authors.initials: B
authors.lastname: August
authors.forename: Sebastian Y
authors.initials: SY
authors.lastname: Bednarek
mesh_terms: D017360:Arabidopsis / Q000378:metabolism / Q000648:ultrastructure*; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient / Q000379:methods; D022163:Clathrin-Coated Vesicles / Q000737:chemistry / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*