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Merck

Isolation and Quantification of Plant Extracellular Vesicles.

Bio-protocol (2017-09-05)
Brian D Rutter, Katie L Rutter, Roger W Innes
ABSTRAKT

Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the apoplastic wash of Arabidopsis thaliana leaves. The isolated EVs can be quantified using a fluorometric dye to assess total membrane content.

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Sigma-Aldrich
Sodium chloride, BioXtra, ≥99.5% (AT)
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MES hydrate, ≥99.5% (titration)
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Murashige and Skoog Basal Salt Mixture (MS), powder, suitable for plant cell culture
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Protease Inhibitor Cocktail, for plant cell and tissue extracts, DMSO solution
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OptiPrep Density Gradient Medium, used for cell and subcellular organelle isolation
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Dipropyl disulfide, 98%
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Trizma® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
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Calcium chloride, anhydrous, granular, ≤7.0 mm, ≥93.0%