I tried to inhibit extracellular microvesicles(MVs) secreted from cells, through using Rock inhibitor(Y27632), calpeptin and several siRNAs(siCDC42, siTSG101, siARF1), but failed to decrease MVs. I've employed a two-step centrifugation process to separate MVs from cell debris. The first centrifugation step at 2,000g removes larger cellular debris, while the subsequent centrifugation at 10,000g isolates the MVs. MVs have been detected using Western blotting, targeting markers such as TSG101, Annexin A1, and Flotillin-2. Additionally, FACS detection using Annexin V has been employed.