Table 3:

Complex II assay flow-chart

REAGENTS	BLANK	SAMPLE (−) Malonate	SAMPLE (+) Malonate
(1) Water	85 μL	95 – x μL	85-x μL
(2) Protein solution	-	X μL	X μL
(3) Reagent Mixture-I	100 μL	100 μL	100 μL
(4) Incubate the plate at 300C for 10 min with a gentle agitation
(5) Reagent Mixture-II with a brief mixing	50 μL	50 μL	50 μL
(6) Add 1M Malonate	10 μL	-	10 μL
(7) Start reaction by adding 2.5 mM DB(8) Measure the absorbance at A600nm for 3–5 min	5 μL	5 μL	5 μL