Journal: Methods in Molecular Biology

Article Title: Lentiviral Vectors and Exosomes as Gene and Protein Delivery Tools

doi: 10.1007/978-1-4939-3753-0

Figure Lengend Snippet: Fig. 1 Exosome purifi cation. ( a ) The experimental workfl ow used for fl uorescent Exo isolation based on differential ultracentrifugation. Exo-containing condi- tioned medium from BODIPY fatty acid-labeled cells is processed by differential centrifugation to remove intact cells and cell debris. Resuspended Exo plus con- taminating proteins pellet is fi ltered using a 0.22 μm membrane fi lter before last centrifugation step. The speed and length of each centrifugation are indicated. The fi nal Exo pellet is resuspended in PBS and can be either directly FC counted or ( b ) further purifi ed by running overnight on an 10–40 % OptiPrep density gra- dient. The fl uorescent peak displays a density ranging from 1.06 to 1.15 g/mL typical of Exo [ 10 , 11 ]. ( c ) Western blot analysis of purifi ed exosomes (5 × 10 7 ) probed with antibodies against Exo markers HSP90, Alix, CD63, and Tsg101

Article Snippet: Nitrocellulose membrane (0.45 μm, Bio-Rad Laboratories) and thick fi lter paper (Bio-Rad Laboratories).

Techniques: Isolation, Labeling, Centrifugation, Membrane, Western Blot