| Bioactivity | Calcium Orange AM is an intracellular calcium reporter. Specific fluorescence can be detected when free calcium binds to Calcium Orange AM (Ex/Em=549/576 nm). Calcium Orange AM does not enter the vacuoles and does not compartmentalize into acidic vesicles[1]. |
| Invitro | Guidelines[2] (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).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. |
| Name | Calcium Orange AM |
| CAS | 172646-19-4 |
| Formula | C59H62N6O21S |
| Molar Mass | 1223.22 |
| Transport | Room temperature in continental US; may vary elsewhere. |
| Storage | Please store the product under the recommended conditions in the Certificate of Analysis. |
| Reference | [1]. Lam CM, et al. Monitoring cytosolic calcium in the dinoflagellate Crypthecodinium cohnii with calcium orange-AM. Plant Cell Physiol. 2005 Jun;46(6):1021-7. [2]. Inami W, et al. Intracellular calcium ion concentration measurement using a phase-modulation fluorescence lifetime method with compensation for phase shift due to the presence of proteins. Anal Sci. 2013;29(2):199-203. |