Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts |
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Authors: | Robert Stéphanie Zouhar Jan Carter Clay Raikhel Natasha |
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Affiliation: | Department of Botany and Plant Sciences, Institute for Integrative Genome Biology, Center for Plant Cell Biology, University of California, Riverside, California 92521, USA. |
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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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