Table 2:

Complex I-III assay flow-chart

REAGENTS	BLANK	SAMPLE (+) Rotenone	SAMPLE (−) Rotenone)
(1) 50 mM KP Buffer	235 μL	232.5 – x μL	235 –x μL
(2)Protein solution	-	X μL	X μL
(3)0.2 mM KCN	2.5 μL	2.5 μL	2.5 μL
(4)4 mM Cyt-c (oxidized)	5 μL	5 μL	5 μL
(5)1.0 mM Rotenone	2.5 μL	-	2.5 μL
(6) START the reaction by adding 5 mM NADH	5 μL	5 μL	5 μL
(7) Measure the absorbance at A550nm for 10 min