27
regarding the blotting results, we can conclude that the suggested
methodology is appropriate for exosome isolation from tissues and
organs.
Finally, the transmission electron microscope (TEM) images in
Fig.  3 for the brain (A)-, heart (B)-, and liver (C)-derived exosomes further demonstrated the success of the ultrasonicationassisted isolation protocol. The exosomes obtained had a diameter
between 50 and 150 nm. The blue arrows seen in the figure clearly
show the stained membranes proving exosomal character.
2 Materials
1. Freshly removed mouse brain, heart and liver. One-mouse hemi
brain was used for exosome isolation.
2. 70% Ethanol.
3. Sulfuric acid (H 2 SO 4 ).
4. Sterile scissors, tweezers/forceps, and razor blade for dissecting
and mincing the organs.
2.1 Reagents Used
for Exosome Isolation
Table 1
Protein content, particle count and RNA content of isolated exosomes
BCA,
O.D.
Protein conc.,
μg.mL
−1
ELISA CD63,
O.D.
Number of CD63+
exosomes
RNA content,
ng.μL
−1
Brain
exosomes
3093
3781
0.195
2.6 × 10
9
45.5
Heart
exosomes
2217
2663
0.099
6.0 × 10
8
6.3
Liver
exosomes
3349
4107
0.305
4.9 × 10
9
143.0
Fig. 2 Western blot analysis of CD63 and CD81 exosomal markers for brain-,
heart- and liver-derived exosomes
Ultrasonics-Assisted Effective Isolation and Characterization of Exosomes from Whole…