The mitochondrial membrane potential was measured in HK2 cells seeded at a density of 105 cells per well on a 24-well plate, as previously described [29 (link)]. Briefly, tetramethylrhodamine, ethyl ester (TMRE; Abcam, Cambridge, MA, USA) (20 nM) was added to the cells and incubated for 30 minutes. After washing three times with 500 ml of phsphate buffered saline/0.2% fetal bovine serum three-times, the cells were read using a FilterMax F3 multimode microplate reader (Molecular Devices, Sunnyvale, CA, USA) at excitation and emission wavelengths of 549 nm and 575 nm, respectively.
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Mitochondrial Membrane Potential Assay in HK2 Cells
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Other organizations : Jeju National University
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Variable analysis
independent variables
- None explicitly mentioned
- Mitochondrial membrane potential
- Cell density (10^5 cells per well)
- Plate type (24-well)
- TMRE concentration (20 nM)
- Incubation time (30 minutes)
- Washing steps (3 times with 500 ml PBS/0.2% FBS)
- Excitation wavelength (549 nm)
- Emission wavelength (575 nm)
- Microplate reader (FilterMax F3)
- No positive or negative controls were explicitly mentioned in the protocol.
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