TMRE accumulates and fluoresces orange in polarized mitochondria. Fluorescence drops dramatically on depolarization in apoptotic or metabolically stressed cells. Analyze using a flow cytometer, fluorescence plate reader, or fluorescence microscopy.
Samples:
Cell culture
Application Notes:
Application Notes: 1. Prepare samples., 2. Create controls with CCCP., 3. Dilute 10X Assay Buffer 1:10 with diH2O., 4. Reconstitute MitoPT TMRM with DMSO., 5. Dilute MitoPT TMRM with 1X Assay Buffer., 6. Add MitoPT TMRM to each sample., 7. Incubate 15-30 minutes., 8. Wash cells: add 1X Assay Buffer and spin cells., 9. Remove supernatant and resuspend cells for analysis., 10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. MitoPT TMRM excites at 548 nm and emits at 573 nm (orange)
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