Table 1:

Complex-I assay flow chart

REAGENTS	BLANK	SAMPLE (−) Rotenone	SAMPLE (+) Rotenone
(1) 50 mM KP Buffer	241 μL	241 μL – x μL	238.5 μL – x μL
(2) Protein solution	-	X μL	X μL
(3) 0.2 M KCN	2.5 μL	2.5 μL	2.5 μL
(4) 5 mM NADH	2.5 μL	2.5 μL	2.5 μL
(5) 1 mM Rotenone	2.5 μL	-	2.5 μL
(6) Read the baseline A340nm for 2 minutes
(7) Start the reaction by adding 1.6 mM decylubiquinone	1.5 μL	1.5 μL	1.5 μL
(8) Read the absorbance at A340nm for 2–3 minutes