Fei tecnai t12 electron microscope

As suggested by the International Society of Extracellular Vesicles (ISEV), a guideline for standardized characterization of particular EVs has been proposed [25 (link)], leading to further reproducible information through different studies. In this study, the protein markers and morphologies of the isolated samples were evaluated. The proteins in EVs were dissolved in RIPA buffer. The solution was boiled at 95 °C for 5 min and then the mixture was spined at 10,000g for 5 min to obtain the clean protein solution. The solution was loaded onto an SDS-PAGE gel (5% stacking gel, 12% running gel, Bio-Rad). After electrophoresis and transferring, the polyvinylidene fluoride (PVEF) membrane (Bio-Rad, Munich, Germany) was incubated with 0.1% BSA in TBST at room temperature for 1 h. The primary antibody was added onto the membrane and incubated at 4 °C for 12 h. After washing, the secondary antibody was incubated with the membrane at room temperature for another 1 h. The membrane was rinsed for three times. Finally, the membrane was detected under the ChemiDoc XRS imaging system (Bio-Rad, Munich, Germany) using an enhanced chemiluminescence (ECL) system (Thermo Scientific, OR, USA). The concentration and size were measured with nanosight (Malvern, UK) and FEI Tecnai™ T12 electron microscope (Thermo Scientific, OR, USA).