Hi,
All the publications using vesicles purification, start with an osmotic lysis.
For exemple this protocol : https://www.nature.com/articles/nprot.2013.053
Here, they use the following homogenization buffer : 320 mM sucrose and 4 mM HEPES (pH 7.4, adjusted with NaOH).
My question is why for this purification, osmotic lysis is more suitable than detergent lysis? If it is.
Does vesicles are not affected by the osmotic lysis like the rest of the cell?
Should we see a difference between those two purifications in western blot analysis of the vesicles?
Respectfully,
Simon.