Exosome Isolation Methods

Isolation of EVs from a plasma by Immunostep SEC columns. Sample obtained can be subsequently analyzed by Abs (280) (Fig. A) and Abs (260/280) (Fig. B), which allows monitoring the elution of the sample, by conventional cytometry with the use of kits based on beads (Fig. C), such as Exostep (Ref. ExoS-25-P81).

A comparison was made using columns competitor, obtaining Immunostep columns a better performance in terms of recovery. A) Comparison of the diameter of the isolated particles by column type. The D50 value means 50% of the particles have that diameter or less. B) Concentration (particles / ml) obtained from purifying plasma (500 µl).

An adequate gating strategy FSC / SSC for 6 micron bead size and PerCP/APC, PerCP-Cy5/ APC or PerCP-Cy5.5/APC helps bead population identification and discrimination of doublets on flow cytometer. Have a look to this dot-plot gating strategy for acquisition and analysis FSC vs SSC (A) and PerCP vs APC (B).