FIGURE 8.

Two centrifugations are crucial to reduce residual platelets. a. ETDA‐ and ACD‐plasmas were collected according to the schematic overview, where our current protocol was compared to the one recommended by the ISTH. b. The number of residual platelets in EDTA‐ and ACD‐plasmas after the two isolation protocols was as measured by a Sysmex automated counter. N = 4. c. The EV‐TRACK knowledgebase was searched for all human plasma studies, and the methods sections of the identified studies were manually read to identify how the blood/plasma was centrifuged prior to freezing and EV isolation. In total, 244 studies were analysed. d. EVs were isolated with Protocol 1 starting with 6‐ml plasma. The presence of the EV markers CD81, CD9, and flotillin‐1, the platelet marker CD41a and the endoplasmic reticulum marker calnexin was determined in SEC fractions 7–14 (36 μl/fraction) by Western blotting. CD41a is detected at 140 kDa, which is the lower band highlighted with *. Platelets pelleted from platelet‐rich‐plasma were used as a positive control for calnexin