Mag-Fura-2 tetrapotassium

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[1].
Monitoring of Ca²⁺ release:
1. Incubate dispersed parotid acinar cells with 8 μM Mag-fura-2 tetrapotassium for 30 min at 37°C. (acinar cells for example) (Mag-Fura-2 tetrapotassium can be introduced into cells using methods such as patch clamping and microinjection).
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.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.