ATP-Red 1

ATP-Red 1 is a multisite-binding switchable fluorescent probe, and can selectively and rapidly responds to intracellular concentrations of ATP in living cells (Ex/Em = 510/590 nm). In Vitro:1. Solution preparation
1.1 Preparation of stock solution
Solvent: DMSO
Concentration: 10 mM is recommended.
Storage: Store at -20°C or -80°C in dark after aliquoting. Avoid repeated freezing and thawing.
1.2 Preparation of working solution
Dilute to 1-10 μM with PBS or serum-free cell culture medium (optimized according to the experiment).
Note: The working solution should be prepared and used immediately. Keep it away from light.

2. Cell staining (suspended cells)
2.1 Collect cells by centrifugation and wash twice with PBS for 5 minutes each time. The cell density is 1×10⁶/mL.
2.2 Add ATP-Red 1 at a final concentration of 5-10 µM.
2.3 Incubate cells at room temperature or 37°C for 15 minutes (or up to 30 minutes for live cell imaging).
2.4 Centrifuge at 400 g for 3-4 minutes and discard the supernatant.
2.5 Add PBS to wash the cells twice, 5 minutes each time.
2.6 Resuspend the cells of serum-free medium or PBS and observe using a fluorescence microscope or flow cytometer.
3. Cell staining (adherent cells)
3.1 Culture the adherent cells on a sterile coverslip.
3.2 Remove the coverslip from the culture medium and remove the excess culture medium. 3.3 Add ATP-Red 1 at a final concentration of 5-10 µM.
3.4 Incubate cells at room temperature or 37°C for 15 minutes (or up to 30 minutes for live cell imaging).
3.5 Aspirate the dye working solution, wash 2-3 times with culture medium, 5 minutes each time, and observe using a fluorescence microscope (Ex/Em = 510/590 nm) or flow cytometer. Note: If flow cytometry is required, the cells need to be digested with trypsin and resuspended before staining.