I followed the protocol of 'abcam', and used FCCP (20uM/well, incubated for 30 mins) as positive control to measure HepG2 mitochondrial membrane potential by TMRE in 96-well plate, but the result of FCCP was even higher than cells untreated.
The cell and TMRE concentration were 10,000/well and 500nM respectively. I added PBS/0.2%BSA after TMRE medium being removed and didn't repeat this step before measured.
Does anyone have ideas about it?
Thanks very much.
Mauro Serricchio
If your signal after FCCP treatment is higher than the untreated cells, your are probably above the quenching concentration of TMRE. This means you should decrease the amount of TMRE you add to the cells. Try 50-100 nM instead of the 500 nM. Good luck
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