MitoMark Red I

Guidelines (The following is a recommended protocol. This protocol is for reference only and needs to be adjusted according to your actual requirements).

1. Stock Solution Preparation
1.1 Prepare a 1 mM stock solution in anhydrous DMSO.

2. Working Solution Preparation
2.1 Dilute with Hepes buffer.
2.2 Working concentration: 100-500 nM.
2.3 Notes: The working solution concentration can be adjusted according to requirements; prepare fresh before use.

3. Staining Procedure
3.1 Cell staining (suspension cells)
3.1.1 Collect cells by centrifugation, add Hepes buffer and wash twice, 5 minutes each time.
3.1.2 Gently resuspend the cells in the staining solution and incubate at 37°C for 30 minutes.
3.1.3 Centrifuge at 400 g for 3-4 minutes and discard the supernatant.
3.1.4 Add Hepes buffer to wash the cells twice, 5 minutes each time.
3.1.5 After resuspending the cells in Hepes buffer, observe using a fluorescence microscope.

3.2. Cell staining (adherent cells)
3.2.1 Culture adherent cells on sterile coverslips.
3.2.2 Remove the coverslip from the culture medium and aspirate excess medium.
3.2.3 Add the dye working solution, rock gently to ensure complete coverage of the cells, and incubate at 37°C for 30 minutes.
3.2.4 Aspirate the dye working solution and replace with 1x Hanks buffer (containing 20 mM HEPES), then observe using a fluorescence microscope.


4. Detection and Analysis
4.1 Instrument type: fluorescence microscope.
4.2 Result analysis: MitoMark Red I specifically labels mitochondria and emits red fluorescence, enabling direct visualization of the relative localization of mitochondria and free radicals within cells.