MitoDNA™ Red 680

Before using MitoDNA™ Red 680 for the first time, allow it to thaw at room temperature. Then, briefly centrifuge it to collect the dried pellet.

1. Prepare cells in a growth medium.

2. Stain cells with MitoDNA™ Red 680 working solution.

3. Incubate samples for 5 to 15 minutes at 37 °C.

4. Monitor fluorescence intensity at Ex/Em = 600/680 nm.

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

1. Prepare a 5 to 10 mM MitoDNA™ Red 680 stock solution in DMSO. For example, add 205 μL of DMSO to the MitoDNA™ Red 680 vial to create a 10 mM stock solution.

   Note: Prepare a single aliquot of the unused MitoDNA™ Red 680 stock solution and store it at ≤ -20 º C, protected from light. Avoid repeated freeze-thaw cycles.

1. Prepare a 5 to 10 μM working solution by diluting the MitoDNA™ Red 680 stock solution in Hanks' solution with 20 mM HEPES buffer (HHBS).

   Note: For optimal results, use this solution within a few hours of preparation.

   Note: Cover the working solution with foil or store it in a dark place to protect it from light.

1. Plate the cells in a 96-well plate with black walls and a clear bottom.

2. Remove the cell culture medium and add 100 µL of MitoDNA™ Red 680 working solution directly to the cells.

3. Incubate the cells at 37°C for 5-15 minutes, protected from light.

   Note: The concentration and incubation time of MitoDNA™ Red 680 may vary depending on the cell line. Test different concentrations to determine the optimal dose.

4. Remove the dye working solution and wash the cells twice with HHBS buffer.

5. Add HHBS buffer and analyze the cells using a fluorescence microscope with excitation/emission settings of 600/680 nm.