To a 5 mL round bottom flask was added 1 mL of DPPC (5 mM in CHCl3), 1 mL of 14:0 PEG2000 PE (0.05 mM in CHCl3) and 1 mL of [1] 2+ (PF6)2 (0.05 mM in acetonitrile). The organic solvents were evaporated under vacuum leading to deposition of lipids on the flask wall. The film was dried under high vacuum for at least one hour and hydrated with phosphate buffer pH of 7.0 and added 15 mg of NADH. The dispersed lipid film was repeatedly freeze-thawing, using liquid N2 and a water bath at 52 °C, forming the giant vesicles. To obtain liposomes of uniform size, the dispersion was extruded at 52 °C through 200 nm cellulose membrane filters 11 times with an Avanti Polar Lipids miniextruder. The liposome mixture was subjected to a Sephadex G-25 size exclusion chromatography (SEC) column (6 cm length, 2 cm diameter) using phosphate buffer pH 7.0 as eluent; the liposome loaded with NADH was collected as the first band.