amicon ultra-15 ultrafiltration tubes  (Merck KGaA)

Journal: Human Cell

Article Title: Blocking secretion of exosomes by GW4869 dampens CD8 + T cell exhaustion and prostate cancer progression

doi: 10.1007/s13577-025-01257-0

Figure Lengend Snippet: Characteristics of PCa exosomes. Human PCa cell lines PC-3 and mouse RM-1 were cultured in exosome-free media for 24 h. Subsequently, exosomes were isolated and purified from the CM of the above cultured cells using ultrafiltration and the phosphatidylserine affinity method. A. Electron microscopy images of exosomes derived from PC-3 or RM-1 cells. B. Particle size analysis of PCa exosomes. After isolation and purification, the diameter of exosomes derived from PCa cells was detected by a nanoparticle size analyzer. C. Analysis of the expression of TSG101, CD9, calnexin, and CD63 on PCa exosomes by western blot. D. CD9 expression on exosome membranes was determined by flow cytometry

Article Snippet: After filtration, 100 mL of the cell supernatant was added to a Millipore ultrafiltration tube (Amicon Ultra-15, Millipore, USA) and centrifuged at 6400 × g for 40 min at 4 °C to collect the crude exosome extract.

Techniques: Cell Culture, Isolation, Purification, Electron Microscopy, Derivative Assay, Particle Size Analysis, Expressing, Western Blot, Flow Cytometry