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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
RESUMO

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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Trizma® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
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MES, ≥99.5% (titration)
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Meio de gradiente de densidade OptiPrep, used for cell and subcellular organelle isolation
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Calcium chloride, anhydrous, granular, ≤7.0 mm, ≥93.0%
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Coquetel Inibidor de Protease, for plant cell and tissue extracts, DMSO solution
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