Purification of Subpopulations of Extracellular Vesicles: A Technique to Purify Subpopulations of Extracellular Vesicles Using Density Gradient Ultracentrifugation
Published: April 30, 2023
Abstract
Source: Urwitz, S. N. et al. Extraction of Extracellular Vesicles from Whole Tissue. J. Vis. Exp. (2019)
This video demonstrates the density gradient ultracentrifugation-based purification method to isolate subpopulations of extracellular vesicles suitable for characterization studies.
Protocol
1. Density Gradient Purification Add 1.5 mL of 60% iodixanol (stock solution in water) to the 1.5 mL sucrose/Tris buffer containing EVs to create a final solution containing 30% iodixanol. Pipette up and down several times to mix solution thoroughly. NOTE: Be cautious to avoid losing solution in the pipette tip, as the high iodixanol concentration is quite viscous. Transfer the 30% iodixanol buffer solution containing EVs to the bottom of a 5.5 mL ultracentri…
Divulgations
The authors have nothing to disclose.
Materials
| 0.45 µm filter | VWR | 28145-505 | |
| 12 mL ultracentrifuge tubes | Beckman Coulter | 331372 | |
| 5.5 mL ultracentrifuge tubes | Beckman Coulter | 344057 | |
| FA-45-6-30 rotor | Eppendorf | 5820715006 | |
| MLS-50 swinging-bucket rotor | Beckman Coulter | 367280 | |
| Optima MAX-XP Benchtop Ultracentrifuge | Beckman Coulter | 393315 | |
| Optima XE-100 ultracentrifuge | Beckman Coulter | A94516 | |
| Optiprep | Sigma | D1556 | 60% iodixanol in sterile water solution |
| Refracto 30PX (refractometer) | Mettler Toledo | 51324650 | |
| S-4-104 rotor | Eppendorf | 5820759003 | |
| SW 41 Ti swinging-bucket Rotor | Beckman Coulter | 333790 | |
| Tabletop 5804R centrifuge | Eppendorf | 22623508 |
Tags
Citer Cet Article
Purification of Subpopulations of Extracellular Vesicles: A Technique to Purify Subpopulations of Extracellular Vesicles Using Density Gradient Ultracentrifugation. J. Vis. Exp. (Pending Publication), e20632, doi: (2023).