. Author manuscript; available in PMC: 2020 Jun 1.

Published in final edited form as: Curr Protoc Toxicol. 2019 Apr 5;80(1):e73. doi: 10.1002/cptx.73

Table 4:

Complex II-III assay flow-chart

REAGENTS	BLANK	SAMPLE (−) Malonate	SAMPLE (+) Malonate

(1) 0.1 M KP buffer, pH 7.4	217 μL	227-x μL	217-x μL
(2) Protein solution	-	X μL	X μL
(3) 50 μM KCN	10 μL	10 μL	10 μL
(4) 0.5M Succinate	10 μL	10 μL	10 μL
(5) Mix and pre-incubate the plate at 30°C for 20 min
(6) 1M Malonate	10 μL	-	10 μL
(7) Start reaction by adding 2.5 mM cytochrome-c (oxidized)	3μL	3μL	3μL
(8) Measure the absorbance at A_550nm for 3–5 minutes