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Thank you for your reply and helpful advices.
I checked the background :
Control well medium +all reagents (including JC-1)
Control well medium+ assay buffer A+B (without JC-1)
Control well medium + only assay buffer A
Control well medium + only assay buffer B
You can find that in medium + all reagents the fluorescent intensity is extremely high compare to other background samples. We think that not all the JC-10 is quenched and this may cause to the high fluorescence reading of the background sample.
Did you get similar results when you checked the kit?
Do you have any other explanation?
You can find attached Excel file containing the results
Asked on May 01 2013
I can confirm that the numbers seems to be alright to me.
The product is used for mitochondria membrane potential measurement.
The dye needs to get into the live cells to exhibit either green (in cytosol) or red (in mitochondria) fluorescence.
Therefore just the solution alone might have some green (monomer) or red (aggregates) fluorescence. This fluorescence has no informational value.
When the mitochondria membrane potential changes, the ratio of the fluorescence in green and red changes. This is the read out the kit provides.
Please read the signal form the bottom of the cell and do not subtract the background signal for the above reasons.
Answered on May 01 2013