Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts
Robert, Stephanie; Zouhar, Jan; Carter, Clay; Raikhel, Natasha
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.
2007, Volume: 2, number: 2, pages: 259-262
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