- Prepare samples.
- Create controls with CCCP.
- Dilute 10X Assay Buffer 1:10 with diH2O.
- Reconstitute MitoPT TMRE with DMSO.
- Dilute MitoPT TMRE with 1X Assay Buffer.
- Add MitoPT TMRE to each sample.
- Incubate 15-30 minutes.
- Wash cells: add 1X Assay Buffer and spin cells.
- Remove supernatant and resuspend cells for analysis.
- Anazlyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. MitoPT TMRE excites at 549 nm and emits at 574 nm (orange).
Rehklau K, Hoffmann L, Gurniak CB, Ott M, Witke W, Scorrano L, Culmsee C, Rust MB. Cofilin1-dependent actin dynamics control DRP1-mediated mitochondrial fission. Cell Death Dis. 2017. Oct 5;8(10):e3063. doi: 10.1038/cddis.2017.448. Abstract
" ... overnight at 4 °C. Mitochondrial membrane potential. To quantify membrane potential, cells. were stained with the potentiometric fluorescent tetramethylrhodamine ethyl ester. (TMRE) dye (MitoPT TMRE kit; Immunochemistry Technologies, Bloomington, MN,. USA).