Exosomes were purified from the conditioned medium (CM) of HepG2 cells using ultracentrifugation methods as described previously.31 Briefly, the CM was collected from HepG2 cells, and then centrifuged at 300 × g for 10 min, 2000 × g for 15 min, and 10,000 × g for 30 min to remove cell debris and additional cellular fragments. After that, the supernatants were ultracentrifuged at 120,000 x g for 70 minutes to isolated exosomes. The pelleted exosomes were then resuspended in 100 μL PBS and stored at −80°C. Nanoparticle tracking analysis (NTA) was used to analyze the quantity and size of exosomes. A JSM-7800F transmission electron microscopy (TEM, StarJoy, Japan) was utilized to identify the morphology of exosomes. The HepG2 cell-derived exosomes were analyzed by Western blotting using the following primary antibodies: anti-TSG101 and anti-CD63.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.