Determination of protein concentration of LEAP‐purified pEVs
LYSED, LYSIS
To determine the amount of protein within each pEV sample, a bicinchoninic acid (BCA) assay was performed following the manufacturer's protocol (Thermo Scientific, Scoresby, Victoria, Australia). Briefly, 20 μL of the sample was homogenized with 60 μL of radioimmunoprecipitation assay buffer (lysis buffer) and incubated at room temperature for 5 min. To each well of a 96‐well plate, 25 μL of lysed sample and 200 μL of working reagent were added, then incubated for 30 min at 37°C before being analysed on the spectrophotometer (Clariostar, BMG Labtech, Germany) at 562 nm.