I am trying to use Fluorescence Spectrophotometer to trace the change in Mitochondrial Membrane Potential. I use potassium chloride based medium to keep the mitochondria energized. I need to use digitonin to permeabilize the cell since I have to add recombinant proteins directly to the cells. Now the problem is:
I have tried various recipe for the KCl based buffer but I am facing the same problem. As soon as I suspend the cells in the buffer they instantly start depolarizing ( as seen by the release of TMRM from the mitochondria and increase in fluorescence.
Can anyone help me to troubleshoot this problem.
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Mitochondrial Membrane Potential Measurement
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