MitoTracker Orange CMTMRos

Guide (The following is our recommended experimental protocol. This protocol is for reference only and should be modified according to your specific needs).
Cell labeling^[1]:
For live cells, follow steps 1-7. For fixed cells, follow steps 8-13.
1. Prepare a labeling solution at a concentration of 25–500 nM in the desired cell culture medium, and warm it to the cell culture temperature.
2. Remove the culture medium from the cells on the coverslip.
3. Immerse the cells in the labeling medium and incubate at 37°C for 15-45 min.
4. Remove the labeling medium and rinse the cells three times with culture medium.
5. Mount the cells.
6. Examine the cells using a fluorescence microscope. If fluorescence staining is too low, try one of the following:
i. Incubate for an additional 30 min in normal culture medium to allow thiol binding to occur.
ii. Increase the initial concentration of the labeling solution.
7. When fluorescence is sufficient, image the cells.
8. Repeat steps 1-4, but use a labeling solution with a working concentration between 100 and 1000 nM.
9. Wash the cells three times with PBS.
10. Fix the cells in PBS (pH 7.4) containing 3.7% formaldehyde for 10 min.
11. Aspirate and wash the cells three times with PBS, 5 min each.
12. Mount with a coverslip.
13. Image the cells.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.