Is there any method to check the complex 1 activity of mitochondria?
Article Assessment of mitochondrial respiratory chain enzymatic acti...
For isolated mitochondria the following paper gives you for all complexes including complex I
Rate limiting step in electron transport: osmotically sensitive diffusion of quinones through voids in the bilayer. J.C. Mathai, Z.E. Sauna, O. John and V. Sitaramam. J. Biol. Chem., 268, 15442-15454, 1993.
Standard complex I activity assay monitors the decrease in the absorbance of NADH (at 340 nm) using decylubiquinone as an artificial electron acceptor. The nature protocol paper is the one I follow. Just a word of caution from my experience: there is considerable rotenone-insensitive background activity, especially in mito from cultured cells.
Good luck!
NADH is impermeable to intact mitochondria. Hence you should use sonicated mitochondrial particles (SMP) for monitoring the decrease in the absorbance of NADH (at 340 nm) using decylubiquinone as an artificial electron acceptor.
The detailed methodology for Complex I activity measurements in mitochondria was described by A.D. Vinogradov in PMID 11124957. Good luck.
Article Dithiothreitol abrogates the effect of arsenic trioxide on n...
pls check my paper
Surendra Katyare Hi, I will be thankful if you can send a detailed protocol for this. in most of the papers, steps are described very vaguely.
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