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P-glycoprotein activity assay

JL Jin Hee Lee
KJ Kyung-Ho Jung
JP Jin Won Park
SM Seung Hwan Moon
YC Young Seok Cho
KL Kyung-Han Lee
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The activity of p-glycoprotein-mediated drug efflux was assessed by Efluxx-ID Green assays (ENZO Life Sciences, Lȍrrach, Germany) following the manufacturer’s protocol50. Briefly, cells harvested by trypsinization were pre-incubated for 5 min at room temperature in PBS containing 2% FBS. The buffer contained 20 μM verapamil, an MDR1 inhibitor, in DMSO or DMSO (vehicle). After Efluxx-ID Green dye was added, the cells were incubated for 30 min at 37 °C. Then, 10,000 cells per sample underwent FACS analysis on a Calibur flow-cytometer using CellQuest software (Becton-Dickinson, NJ). Efluxx-ID Green was excited at 490 nm, and fluorescent emissions were detected at 514 nm.

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