Calcium Orange AM

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[2].
1. Calcium Orange AM is freshly prepared in dehydrated DMSO before each experiment.
2. Prepare the recording medium for measuring [Ca²⁺]i in cells consisted of 20 mM HEPES, 115 mM NaCl, 5.4 mM KCl, 0.8 mM MgCl₂, 1.8 mM CaCl₂, and 13.8 mM D-glucose adjusted to pH 7.4 by adding 1 M KOH.
3. Exchange the culture medium for 1 mL of a recording medium including 10 μM Calcium Orange AM.
4. Incubate cells under conditions of 5% CO₂ and 37°C for 40 min.
5. Remove the recording medium, including calcium orange AM from the dish, and add the culture medium after washing it three times with 1 mL of the recording medium.
6. Incubate the cells for 1 h. After incubation, exchange the culture medium for the recording medium again.
7. Detect fluorescence.

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