fig2

Figure 2. Free flow electrophoresis (FEE) effectively separates components of MSC-EV preparations. (A) A five-step pH-profile (pH 4.8, 5.4, 6.4, 7.4, and 8.4) was used for the fractioning of a 133 μL sample of a well-characterized MSC-EV preparation that was applied at the vertical position corresponding to that of the collected Fraction 78. An interval zone electrophoresis was performed at 1000 V for 6 min in a 6 cm broad separation area. (B) A pherogram of the protein content within the different fractions. (C) The results of dot blot analysis of all 96 obtained fractions using a mixture of anti-CD9 (VJ1) and anti-Syntenin (EPR8102) antibodies. As a control, at Position 96, 15 μg of a PEG precipitated MSC-EV sample were applied. Fractions delivering positive dot blot signals (Fractions 35-37) are labeled by arrows and the black line in (C).