Oct 28, 2024

Public workspaceEV Isolation from Plasma

DOI
dx.doi.org/10.17504/protocols.io.261ge5qywg47/v1
  • Aoife Gahlawat1
  • 1DKFZ
Aoife Gahlawat
Icon indicating open access to content
QR code linking to this content
DOI: dx.doi.org/10.17504/protocols.io.261ge5qywg47/v1
Protocol Citation: Aoife Gahlawat 2024. EV Isolation from Plasma. protocols.io https://dx.doi.org/10.17504/protocols.io.261ge5qywg47/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: October 22, 2024
Last Modified: October 28, 2024
Protocol Integer ID: 110554
Abstract
A protocol to isolate extracellular vesicles (EVs) from human plasma
SEC
SEC
Thaw plasma to Temperature20 °C , you will need Amount500 µL Prepare IZON column 70nm qEV Original by washing with filtered PBS

Once column is washed, add the plasma sample and let it enter the resin before topping up with Amount2 mL
filtered PBS. Collect the eluate by gravity in a falcon and discard as void. Add Amount2.5 mL filtered PBS and collect directly into a 4ml Amicon filter tube with 100kDa cut-off. This is your EV fraction. Either wash the column or repeat to collect a non-EV (protein fraction).

Centrifuge the Amicon tube at 2000 G in 30 second steps until the desired volume is reached - usually Amount300 µL

Qubit Protein Concentration
Qubit Protein Concentration
As an initial QC, protein concentration can be measured by Qubit.