Mag-Fura-2 tetrapotassium

Mag-Fura-2 tetrapotassium is a UV excitable rational fluorescent Mg2+/Ca2+ indicator (Ex=334-360 nm, Em=510 nm). Mag-Fura-2 tetrapotassium can be used for the determination of Mg2+ and Ca2+ concentrations[1][2].

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Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). Monitoring of Ca2+ release[1]: 1. Incubate dispersed parotid acinar cells with 8 µM Mag-fura-2 tetrapotassium for 30 min at 37℃. (acinar cells for example). 2. Wash cells twice with fresh HBSS-H without BSA. 3. Precoate cell adhesive Cell-Tak on sample chambers. 4. Transfer the dye-loaded cells to the chambers and attach to the bottom. 5. Mounte the sample chambers on the stage of an inverted microscope (equipped with a 40 × objective), wash with BSA-free HBSS-H and then with Mg2+/ATP-free ICM. 6. Incubate acinar cells with Mg2+/ATP-free ICM containing 50 µg/mL saponin for 3-5 min at room temperature. 7. Wash the cells with ICM containing Mg2+ and ATP, and incubate in the complete ICM for at least 5 min to allow complete filling of the intracellular Ca2+ stores. 8. Alternately excite permeabilised cells, capture and digitise fluorescence emission at 510 nm by a digital imaging system (record the 344 nm/360 nm ratio every 20 s). Note: ICM (intracellular-like medium) containing 125 mM KCl, 19 mM NaCl, 10 mM HEPES (pH 7.3 with KOH), 3 mM ATP, 1.4 mM MgCl2, 0.33 mM CaCl2, and 1 mM EGTA.

[1]. Tojyo Y, et al. Monitoring of Ca2+ release from intracellular stores in permeabilized rat parotid acinar cells using the fluorescent indicators Mag-fura-2 and calcium green C18. Biochem Biophys Res Commun. 1997 Nov 7;240(1):189-95.

[2]. Dai LJ, et al. Intracellular Mg2+ and magnesium depletion in isolated renal thick ascending limb cells. J Clin Invest. 1991 Oct;88(4):1255-64.