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. Author manuscript; available in PMC: 2019 Apr 22.
Published in final edited form as: Methods Mol Biol. 2018;1740:69–83. doi: 10.1007/978-1-4939-7652-2_7

Figure 2. Schematic illustration of C-DGUC for exosome isolation.

Figure 2.

(1) Place a 2 mL solution of 60% iodixanol below the pre-cleared conditioned medium that serves to cushion the nanoparticles during centrifugation; (2) sediment the nanoparticles at 100,000 × g for 3 hrs; (3) remove the 2 mL cushion along with 1 mL of overlaying medium and place it below a pre-formed step density gradient composed of 3 layers of iodixanol solution, 3 mL each; (4) perform density gradient ultracentrifugation at 100,000 × g for a period of 18 hrs; (5) collect the medium from the top of the tube in 1 mL increments. Exosomes derived from cultured primary BMDM typically are isolated in fractions 6 & 7.