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Mitochondrial membrane potential (ΔΨm) assay

SC Su-Jung Choi
CA Chi-Hyun Ahn
KH Kyoung-Ok Hong
JK Ji-Hoon Kim
SH Seong-Doo Hong
JS Ji-Ae Shin
SC Sung-Dae Cho
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ΔΨm was measured by flow cytometry using the lipophilic fluorescent dye JC-1. Cells were harvested via trypsinization, washed with PBS, and pelleted via centrifugation at 3500 rpm for 5 min. The pellets were resuspended in a 1× JC-1 working solution and incubated at 37 °C for 30 min in the dark. The stained cells were washed with a 1× assay buffer and pelleted via centrifugation at 3500 rpm for 5 min. After removing the supernatant, the cells were resuspended in a 1× assay buffer. Subsequently, the cells were transferred to FACS tubes and analyzed with the FACSCalibur Flow Cytometer. The proportion of stained cells was quantified from 10,000 cells per sample with BD CellQuest™ Pro software. The data were reanalyzed using FlowJo software.

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