SunRed™ Acetate

Protocol summary

1. Prepare cells with test compounds
2. Remove the medium
3. Add SunRed™ Acetate working solution (100 µL/well/96-well plate or 25 µL/well/384-well plate)
4. Incubate at 37°C, 5% CO₂ incubator for 1 hour
5. Wash and replace the working solution with HHBS
6. Read fluorescence intensity at Ex/Em = 620/660 nm (cut off 640 nm)

Important notes
The following is the recommended protocol. It only provides a guideline, should be modified according to the specific needs.

1. SunRed™ acetate stock solution (2 to 10 mM)
Prepare a 2 to 10 mM stock solution of SunRed™ acetate in DMSO. The stock solution should be used promptly.

SunRed™ acetate working solution:
Prepare SunRed™ acetate working solution at 5 to 10 µM in 1X Hank’s salt solution with 20 mM Hepes buffer (HHBS) or buffer of your choice before the experiment.

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

1. Remove the medium from the cells.
   
   
2. Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of SunRed™ acetate working solution.
   
   
3. Incubate the SunRed™ acetate working solution plate at room temperature or 37°C for 1 hour, protected from light. The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment. For non-adherent cells, it is recommended to centrifuge cell plates at 800 rpm for 2 minutes with brake off after incubation.
   
   
4. Monitor the fluorescence intensity at Ex/Em = 620/660 nm (cut off 640 nm).