I tried to isolate exosomes from mouse plasma by ultracentrifugation. I was thinking about how many mice I will need to get a usable concentration of exosomes for FACs analysis as well as T-cell assays. At first I killed 14 mice were I got 6 ml plasma from. After ultracentrifugation I respuspended the exosome pellet in 150 µl PBS and had a concentration of 76 µg/ml. This means that I will need a loooot of mice to get a good concetration like 1-2 mg/ml. Has anyone done that before so far?
Hi Bettina, get in touch with [email protected] He might be able to assist in terms of plasma as a starting material. kind regards ketil
Thank you David for the nice protocol. I have another question for you: "Significant issues arise when using thawed/frozen blood" Is it possible when you do plasma isolation an freeze the sample afterwards that you have really small structures (I think it could be crystal structures) less than 20 nm in electrom microscopy analysis? Because I had this problem when I used -80 thrawed frozen plasma exosomes in comparison with fresh prepared plasma exosomes.But both isolation have been done on ice.
we routinely recover exosomes from small volumes of serum/plasma- mouse and human- typically 100 uL, with Total exosome isolation reagents. even smaller vol can be used- no problem
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