Materials and Reagents

1. Vacuum driven disposable filtration system (Stericup 0.22 μm) (Merck Millipore Corporation, catalog number: P25628X1 )
   
2. Graduated cylinder (50 ml)
   
3. Sephadex G-25 columns (PD-10) (GE Healthcare, catalog number: 17-0851-01 )
   
4. Glass rod
   
5. Thick wall polycarbonate plastic tubes for ultracentrifugation (Beckman Coulter, catalog number: 355631 )
   
6. Glass disposable micro pipet (Corning Inc., catalog number: 7099S-20 )
   
7. Resource Q anion exchange column 6 ml (GE Healthcare, catalog number: 17-1179-01 )
   
8. Chlamydomonas reinhardtii cells (about 8 g of wet weight collected from 2-3 liters of a 1.2 x 10⁷ cells/ml photo-heterotrophic culture)
   
9. Tris (2-amino, 2-hydroxymethyl, 1, 3-propanediol) (Trizma base) (Sigma-Aldrich, catalog number: T-1503 )
   
10. Magnesium sulfate heptahydrate (MgSO₄^.7H₂O) (vidraFOC, Panreac, catalog number: 131404-1210 )
    
11. Magnesium chloride hexahydrate (MgCl₂^.6H₂O) (Merck Millipore Corporation, catalog number: 1.05833 )
    
12. Sodium chloride (NaCl) (AppliChem GmbH, Panreac, catalog number: 121659.1211 )
    
13. Sodium bicarbonate (NaHCO₃) (Merck Millipore Corporation, catalog number: 1.06329 )
    
14. 2-mercaptoethanol (Merck Millipore Corporation, catalog number: 8.05740 )
    
15. Sulfuric acid (H₂SO₄) (Sigma-Aldrich, Fluka, catalog number: 84718 )
    
16. Hydrochloric acid (HCl) (VWR International, J.T.Baker^®, catalog number: 6081 )
    
17. Sucrose (Sigma-Aldrich, Fluka, catalog number: 84100 )
    
18. cOmplete (Protease inhibitors tablets) (Roche Diagnostics, catalog number: 11697 498001 )
    
19. Poly(vinylpolypyrrolidone) (Sigma-Aldrich, catalog number: P-6755 )
    
20. Ammonium sulfate [(NH₄)₂SO₄] (AppliChem GmbH, Panreac, catalog number: 141140 )
    
21. dH₂O (deionized water) (processed by the Milli-Q system from Merck Millipore Corporation)
    
22. Coomassie Blue-stained SDS-PAGE (optional)
    
23. Extraction buffer (EB) (see Recipes)
    
24. Sucrose gradient buffer (SGB) (see Recipes)
    
25. Sucrose density gradient (see Recipes)
    
26. Chromatography buffer A (CBA) (see Recipes)
    
27. Chromatography buffer B (CBB) (see Recipes)
    
28. Activation buffer (AB) (see Recipes)
    

Equipment

1. Ultrasonic processor (Sonics Vibracell, model: VCX 500 ) equipped with a 19 mm high-gain probe
   
2. Optical microscope (Microscope Central, Nikon, model: Alphaphot 2 YS2 )
   
3. Magnetic stirrer (Bibby Scientific, Stuart, model: SB161-3 ) and magnetic bars
   
4. Preparative centrifuge (GMI, Beckman Coulter, model: J2-HS ) equipped with a fixed-angle rotor (Beckman Coulter, model: JA-20 )
   
5. Ultracentrifuge (GMI, Beckman Coulter, model: L-70 ) equipped with a fixed-angle rotor (Beckman Coulter, model: 55.2 Ti )
   
6. Two-chamber (2 x 8 ml) gradient mixer (homemade)
   
7. UV Monitor (GE Healthcare, Amersham-Pharmacia Biotech, model: UV-1 with Rec112 register )
   
8. Fast Performance Liquid Chromatography equipment (FPLC) (GE Healthcare, Amersham-Pharmacia Biotech, model: Äkta Prime )
   
9. Peristaltic pump (GE Healthcare, Pharmacia, model: P-1 )
   
10. UV-V spectrophotometer (Shimadzu Scientific Instruments, model: UV-1603 )
    
11. Water jet vacuum pump (aspirator) (KARTELL SPA VIA DELLE INDUSTRIE, model: 1395 )
    
12. Ultrasonic water bath (Scientific Support, Branson, model: 2200 )
    

Procedure