Protocol for the Analysis of Yeast and Human Mitochondrial Respiratory Chain Complexes and Supercomplexes by Blue Native Electrophoresis

. 2020 Sep 3;1(2):100089. doi: 10.1016/j.xpro.2020.100089

CV Activity Solution

Reagent	Final Concentration	Stock Concentration	Add to 10 mL
Tris base	35 mM	n/a	42.4 mg
Glycine	270 mM	n/a	0.2 g
Magnesium sulfate	14 mM	n/a	16.85 mg
ATP	5 mM	n/a	25.36 mg
Lead nitrate	0.2 % (w/v)	n/a	20 mg

Prepare right before use. Prepare the reaction with Tris, glycine, magnesium sulfate, and ATP in this order. Adjust pH to 7.8 and, then, add lead nitrate to the solution. Adjust pH to 8.6. The solution appears cloudy. Use it freshly prepared, and discard any unused solution.