Hello,
I'd like to isolate exosomes from cell culture supernatant.
but before I need to filter it. I'm familiar with the necessary pore size of the filter I'd be interested in the material of the hydrophilic filter in order to be able to obtain exosomes. Does anyone have experience with cellulose acetate membrane? or which one would you recommend (PES, nylon etc)?
Thanks!
Hello Katalin,
I do filter and concentrate cell culture supernatant on routine basis for protein isolation. In my case Polyethersulphone (PES) membrane works very fine as they retain maximum proteins with greater than 90% efficiency. I have done further proteomic analysis and got good bands of proteins in culture supernatant after filtration and concentration.
PES are quiet good and reliable for such preparations.
Hope it helps !
Filtration will result in loss of exosomes. I ll suggest you to go for differential centrifugation.
Hi Glasenhardt,
If you want to have a pure population of exosomes, I would suggest a combination of differential centrifugation and filtration method. For this-
1. centrifuge the samples at 30,000 g for 30 minutes to remove the cellular debris
2. do a filtration step with 0.22 um syringe filter (nylon) which will effectively remove microvesicles
3. then follow the ultracentrifugation procedure.
Hope it will work.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology