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If there are exosomes and virus in cell culture media, is it possible to isolate only exosomes?

asked Feb 20, 2014 in ExosomesTalk by Caropressman

2 Answers

pretty challenging, as viruses are in the same size range as exosomes and have very similar composition. So give it a try with the Total reagent, but viruses could be co-precipitating to some extent. A better approach would be to use the Total reagent first, and then use Dynabeads with Ab specific to some marker unique to exosomes (to purify them nicely) or on the contrary viral particles (to deplete them)
answered Feb 27, 2014 by Vlassosv

Isolation of subpopulations of exosomes:

If one wants to obtain ultra-pure exosomes – Dynabeads (magnetic beads) decorated with CD9, CD63, CD81 or EpCAM antibodies, are currently available. This will allow to recover very clean population of exosomes, following initial purification e.g with Total Exosome Isolation (from cell media) or ultracentrifugation. These subpopulations of exosomes can be analyzed by electron microscopy, flow cytomtery, WB and RNA contect etc. If this is of interest I may supply a protocol for this for downstream WB or Flow Cytometry.

We have also launched 3 products for isolation of exosome subtypes

  • Exosome – Human CD9 Isolation (Cat. No. 10614D)
  • Exosome – Human CD81 Isolation (Cat. No. 10616D)
  • Exosome – Human EpCAM Isolation (Cat. No. 10618D)

Exosomes isolated in this way are suitable for downstream Western Blotting

I would also like to mention that these Dynabeads mentioned above also works very well in immunoprecipitation. After pre-enrichment the exosomes are lysed. Then one can use either of these beads to pull out e.g. CD81 protein from the lysate and prepare the IP for SDS/Page and western blotting.

If the downstream application is flow detection of exosome subtypes 3 new products have been launched for this as well. They are suitable for both checking the status of your pre-enriched exosomes before doing experiments or in depth flow analysis on the exosome subtypes.

  • Exosome – Human CD9 Flow Detection (Cat. No. 10620D)
  • Exosome – Human  CD81 Flow Detection (Cat. No 10622D)
  • Exosome – Human EpCAM Flow Detection (Cat. No. 10624D)

 

answered Dec 17, 2014 by ketil.pedersen
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