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Protein fordøjelse, ultrafiltrering og størrelse udstødelse kromatografi at optimere Isolation af Exosomes fra humant blod Plasma og Serum
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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
Vi præsenterer her, en protokol for at rense exosomes fra både plasma og serum med reduceret Co rensning af ikke-exosomal blodproteiner. Den optimerede protokol omfatter ultrafiltrering, protease behandling og størrelse udstødelse kromatografi. Forbedret rensning af exosomes fordele downstream analyser, herunder mere nøjagtig kvantificering af vesikler og proteom karakterisering.
