Greetings,

Yes you can, however you will have to mix it with some highly soluble compound, which should also add high density to your buffer. The technique principle is based on the density differences that exist among different layers of a buffer containing different concentration ratio of an osmolyte. I've used sucrose gradient to isolate VLPs, however, as they tend to be kind of capricious with buffer composition, I had to use a mix of sucrose and my refolding buffer, containing TrisHcl and what not. Notwithstanding, if you dont feel like using sucrose, you can always use cessium chloride or, even better, iodixanol.

Best regards

Dear Adán thank you so much for your answer .. Actually I am making the density gradient with Iodixanol and regarding the protocol it should be mixed with sucrose 25mM to obtain the different layers ... the problem is that I am affraid about the possible effect of sucrose on EV . That's I was wondering if I could substitute it with PBS, but I didn't find any protocol mixing Iodixanol with PBS.

Well, as long as every single step has the desired iodixanol concentration, there should be no problem. By the way, AAV standard protocol uses PBS-MK to perform density gradient.

https://www.addgene.org/protocols/aav-purification-iodixanol-gradient-ultracentrifugation/

Good luck

Thank you so much Adán for the protocol that is very helpful ...i 'll try it

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