title: Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts.
abstract: Vacuoles are very prominent compartments within plant cells, and understanding of their function relies on knowledge of their content. Here, we present a simple vacuole purification protocol that was successfully used for large-scale isolation of vacuoles, free of significant contamination from other endomembrane compartments. This method is based on osmotic and thermal disruption of mesophyl-derived Arabidopsis protoplasts, followed by a density gradient fractionation of the cellular content. The whole procedure, including protoplast isolation, takes approximately 6 h.
authors.affiliation: Department of Botany and Plant Sciences, Institute for Integrative Genome Biology, Center for Plant Cell Biology, University of California, Riverside, California 92521, USA.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Robert
authors.forename: Jan
authors.initials: J
authors.lastname: Zouhar
authors.forename: Clay
authors.initials: C
authors.lastname: Carter
authors.forename: Natasha
authors.initials: N
authors.lastname: Raikhel
mesh_terms: D017360:Arabidopsis / Q000737:chemistry*; D002458:Cell Fractionation / Q000379:methods*; D018515:Plant Leaves / Q000166:cytology*; D011523:Protoplasts / Q000737:chemistry*; D014617:Vacuoles / Q000737:chemistry*