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Updated: Jul 15, 2026

Isolation of Protoplasts from Tissues of 14-day-old Seedlings of Arabidopsis thaliana
Published on: August 17, 2009
Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts
Stéphanie Robert1, Jan Zouhar, Clay Carter
1Department of Botany and Plant Sciences, Institute for Integrative Genome Biology, Center for Plant Cell Biology, University of California, Riverside, California 92521, USA.
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.
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