I fear that exosomes would not be in the medium, but in the same density as adipocytes.
Absolutely! To isolate EVs from suspension/non-adherent cells you will first need to pellet the cells in suspension by centrifugation. Anywhere between 500xg-3000xg should do the trick for 10-15 mins; you need to find what protocol works best for your samples!
After low speed centrifugation the EVs will remain in the supernatant/clarified conditioned media. You can discard the cell pellet and then simply process the supernatant according to whatever isolation protocol you are using.
Hope this helped!
You should not have any problem if cells are pelleted first and then follow an ultracentrifugation protocol to isolate exosomes. As Braulio mentions, given the number of protocols available for exosome isolation, better to try 2 or 3 and check enrichment in exosomal markers. Ultracentrifugation worked perfectly in my hands. Good luck.
How would you pellet adipocytes? Do you recommend a centrifugation speed and time?
Hello, Can anyone send me the protocol used routinely to isolate EBV virus from supernatant
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