4.6. Characterization of Small Extracellular Vesicles with Coomassie Assay
TECAN
Coomassie assay was used for detection of protein amounts in small EVs. As standard, a serial dilution of 25 μg/mL to 2.5 μg/mL bovine serum albumin (A3608, Sigma-Aldrich, St. Luis, MO, USA) in PBS–/– was used. Each 30 μL of sample/standard and Coomassie assay reagent (23200, Thermo Fisher Scientific, Waltham, MA, USA) were added to wells in a 384 well plate (781061, Greiner Bio-One, Kremsmuenster, AUT) and gently mixed for 30 s at a plate shaker. After an incubation period of 10 min at room temperature in dark conditions, absorbance was measured at 595 nm using a TECAN SPARK 20M (TECAN, Morrisville, NC, USA). AutoOptical density values of samples were compared to the standard curve generated with known concentrations of protein. 