Hello, We want to perform Cytochrome c oxidase assay to check electron transport chain function in our treated cells. Is it necessary to isolate mitochondria for the assay if yes why?
Further, we can do it with oxygen consumption assay. Can we do this assay with intact cells?
Why not measure the enzyme's activity spectrophotometrically like in attached the procedure? Mitochondria do not need to be isolated but you do need to disrupt/homogenize the cells or tissue.
Please check the publication in the attachment.
Cheers, Christian
Article Assessment of mitochondrial respiratory chain enzymatic acti...
Please see the attached PDF which tells you details of assay using Oxygen electrode. You can monitor the activity in both homogenate and mitochondria.
S. S. Katyare
Professor of Biochemistry
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