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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
摘要

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 ® 碱, Primary Standard and Buffer, ≥99.9% (titration), crystalline
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MES 水合物, ≥99.5% (titration)
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OptiPrep密度梯度培养基, used for cell and subcellular organelle isolation
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氯化钙, anhydrous, granular, ≤7.0 mm, ≥93.0%
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Murashige & Skoog 基础盐混合物 (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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氯化钠, BioXtra, ≥99.5% (AT)
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二丙基二硫, 98%