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Vertering van eiwitten, ultrafiltratie en Size Exclusion Chromatography voor het optimaliseren van het isolement van Exosomes uit menselijk bloed Plasma en Serum
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생화학
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JoVE 신문 생화학
Protein Digestion, Ultrafiltration, and Size Exclusion Chromatography to Optimize the Isolation of Exosomes from Human Blood Plasma and Serum
DOI:

09:22 min

April 13, 2018

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Chapters

  • 00:04Title
  • 01:06Preparation of Crude Sample (Either Plasma or Serum) by Pelleting Larger Vesicles and Cellular Debris
  • 02:04Digestion of Non-exosomal Blood Proteins and Removal of Small Proteins/Peptides by Centrifuge Ultrafiltration
  • 04:21Purification of Vesicles by Size Exclusion Chromatography (SEC)
  • 05:45Concentration of Purified Exosomes/Total Protein Quantification and Quantification/Sizing of Exosomes by Nanoparticle Tracking Analysis (NTA)
  • 07:09Results: Increased Vesicle Yield and Exosome-specific Markers After Purification
  • 08:15Conclusion

Summary

자동 번역

Hier presenteren we een protocol om te zuiveren van exosomes van plasma en serum met verminderde CO zuivering van niet-exosomal bloedeiwitten. Het geoptimaliseerde protocol bevat ultrafiltratie, protease behandeling en grootte uitsluiting chromatografie. Verbeterde zuivering van exosomes voordelen downstream analyses, met inbegrip van nauwkeuriger kwantificering van blaasjes en proteomic karakterisering.

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