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Purification of High Yield Extracellular Vesicle Preparations Away from Virus
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JoVE 杂志 免疫与感染
Purification of High Yield Extracellular Vesicle Preparations Away from Virus

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

DOI:
10.3791/59876-v

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00:07 min

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September 12, 2019

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, , , , , , , ,
1Laboratory of Molecular Virology, School of Systems Biology,George Mason University, 2American Type Culture Collection (ATCC), 3ATCC Cell Systems, 4Ceres Nanosciences, Inc, 5Department of Immunology and Nano-medicine, Herbert Wertheim College of Medicine,Florida International University

Chapters

  • 00:04Title
  • 01:00Filtration and Precipitation of Extracellular Vesicles
  • 02:25Construction of a Density Gradient and Enrichment of EV Fractions
  • 04:06Preparation of Nanoparticle Pellet for Downstream Assay
  • 05:05Results: Increased EV Yield and Characterization of EVs
  • 06:09Conclusion

Summary

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This protocol isolates extracellular vesicles (EVs) away from virions with high efficiency and yield by incorporating EV precipitation, density gradient ultracentrifugation, and particle capture to allow for a streamlined workflow and a reduction of starting volume requirements, resulting in reproducible preparations for use in all EV research.

Tags

Extracellular VesiclesEVsVirus-free EV PreparationsEV PurificationUltracentrifugationPEG PrecipitationIodixanol Density GradientNanoparticle Enrichment

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