Mild Denaturing Electrophoresis

Etioplast/chloroplast membranes were solubilized in a 10 mM Tricine buffer (pH 7.6) containing 20% (w/v) sucrose to a chlorophyll concentration of 0.5 mg mL⁻¹. Next, the n-dodecyl-β-d-maltopyranoside and lithium dodecylsulfate were added to a final concentration of 0.5% (w/v) and 0.05% (w/v), respectively. After 20 min of dark incubation on ice, samples containing 4 μg of chlorophyll were loaded into wells of an SDS-depleted 3% (w/v) polyacrylamide stacking gel. Electrophoresis was performed using a SDS-depleted 8% (w/v) polyacrylamide resolving gel supplemented with 10% (w/v) sucrose. The electrophoresis was conducted with a 25 mM Tris buffer (pH 8.3) containing 192 mM glycine and 0.1% (w/v) lithium dodecylsulfate in a MiniProtean3 electrophoresis cell (Bio-Rad Laboratories).