Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts.
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.
Robert
Stéphanie
S
Department of Botany and Plant Sciences, Institute for Integrative Genome Biology, Center for Plant Cell Biology, University of California, Riverside, California 92521, USA.
Zouhar
Jan
J
Carter
Clay
C
Raikhel
Natasha
N
D017360:Arabidopsis / Q000737:chemistry*; D002458:Cell Fractionation / Q000379:methods*; D018515:Plant Leaves / Q000166:cytology*; D011523:Protoplasts / Q000737:chemistry*; D014617:Vacuoles / Q000737:chemistry*