Comparison of various methodologies for exosome isolation. (a) Schematic representation of exosomes isolation methods. MiaPaCa cells (3 × 106) were cultured in regular media, after 24 h, media was replaced with 5% exosome depleted FBS. After 48 h condition media was collected, centrifuged at 300 × g for 10 min to remove cells and cell-debris. Thereafter, exosomes were isolated using ultracentrifugation and four commercial kits following the manufacturer’s instructions. (b) Levels of proteins were estimated in intact exosomes by DC protein assay. Highest exosome yield was detected for Invitrogen Kit, while MagCapture yielded the least amount of exosomes. Data presented as mean ± s.d.; n = 3, p-values **<0.0093, and ****<0.0001.