Sep 15, 2022
Plasma Exosome Isolation
- Silvia Cerri1
- 11Laboratory of Functional Neurochemistry, IRCCS Mondino Foundation, Pavia, Italy
Protocol Citation: Silvia Cerri 2022. Plasma Exosome Isolation. protocols.io https://dx.doi.org/10.17504/protocols.io.81wgb7xnqvpk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 02, 2020
Last Modified: May 31, 2024
Protocol Integer ID: 45157
Keywords: exosome, isolation, plasma, EV, extracellular vesicles, ASAPCRN
Abstract
This protocol details methods for the isolation of small extracellular vesicles from plasma.
Materials
Materials
- 13mm sterile syringe filters with a 0.8 µm Supor (PES) membrane
- 13mm sterile syringe filters with 0.2 mm Supor (PES) membrane
- 1X PBS
Equipment
- Ultracentrifuge
Safety warnings
Please refer to the Safety Data Sheets (SDS) for health and environmental hazards.
Before start
Keep samples On ice during the entire procedure!
Plasma Exosome Isolation
Plasma Exosome Isolation
2h 30m
2h 30m
Incubate plasma samples (starting volume: 1-1.5ml) On ice for complete thaw.
Filter samples through a 13mm sterile syringe filters with a 0.8 µm Supor (PES) membrane.
Centrifuge at 20000 x g, 4°C, 01:00:00 to remove large extracellular vesicles.
Collect the supernatant and filter it through 13mm sterile syringe filters with 0.2 mm Supor (PES) membrane.
Centrifuge at 100000 x g, 4°C, 01:00:00 to pellet small extracellular vesicles.
Remove the supernatant, resuspend the pellet in 1 mL 1X PBS and repeat the previous step (repeated again in the next step for convenience).
Note
This step is optional, according the final application of the pellet.
Centrifuge at 100000 x g, 4°C, 01:00:00 to pellet small extracellular vesicles.
Process the pellet according to the type of analysis.