Why adding CCCP giving more red fluorescence than green compared to untreated?

Sulagna Chatterjee @Sulagna-Chatterjee-4

17 March 2023 1 3K Report

I am using CCCP (5OuM) for JC1 assay as a positive control but I am getting more red fluorescence than green fluorescence that is no shift from red to green shift compared to untreated cells . What is the solution please suggest.

Tanoy Dutta

Hi Sulagna Chatterjee,

The concentration of CCCP seems fine. However, for mitochondrial membrane potential uncoupling, 20 uM CCCP should be good enough if incubated for 30 mins. How much is the JC-1 concentration in your experiment? I recommend using 10 uM JC-1 dye and incubate for 10 - 15 mins. I guess the emission channels are set at 510 – 540 nm (monomer emission) and 575 – 620 nm (aggregate emission).

All the best.

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