Materials and Reagents

1. Burkitt’s Lymphoma B cell lines (e.g. Mutu^-, Mutu I, Mutu III cell lines)
   
2. RPMI 1640 medium (Wako Chemicals USA, catalog number: 189-02025 )
   
3. Sucrose (Sigma-Aldrich, catalog number: S7903 )
   
4. Anti-CD63 monoclonal antibody (clone MEM-250) (Abnova, catalog number: MAB0931 )
   
5. Bradford protein assay kit (Bio-Rad Laboratories, catalog number: 500-0006JA )
   
6. 1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine perchlorate (DiI) (Life Technologies, catalog number: D3911 )
   
7. Fetal Bovine Serum (FBS) (Sigma-Aldrich, catalog number: F9423 )
   
8. Tris
   
9. NaCl
   
10. EDTA
    
11. Exosome-depleted FBS (see Recipes)
    
12. TNE buffer (see Recipes)
    
13. 0.25-2.5 M sucrose gradient in TNE buffer (see Recipes)
    

Equipment

1. 10 cm dish
   
2. Centrifuge (Eppendorf, model: 5810R or that with equivalent equipment spec)
   
3. Ultracentrifuge (Beckman Coulter, model: Optima L-80 XP or that with equivalent equipment spec)
   
4. 37 °C, 5% CO₂ cell culture incubator
   
5. Autopipette
   
6. 50 ml polypropylene concal plastic tubes (BD Biosciences, Falcon^®, catalog number: 352070 or that with equivalent spec)
   
7. SW28 rotor (Beckman Coulter, model: 342204 )
   
8. SW40Ti rotor (Beckman Coulter, model: 331301 )
   
9. Polyallomer centrifuge tubes 1 x 3½ in (25 x 89 mm) for SW28 rotor (Beckman Coulter, catalog number: 326823 )
   
10. Polyallomer centrifuge tubes 9/16 x 3½ in (14 x 89 mm) for SW41Ti rotor (Beckman Coulter, catalog number: 331372 )
    
11. Spectrometer
    
12. Fluorescent or confocal laser scanning microscope
    

Procedure