Mag-Fluo-4 AM 

Mag-Fluo-4 AM is a fluorecent Ca²⁺ chelator, with high affinity for calcium. Mag-Fluo-4 AM can specifically identify intracellular calcium ions, with high sensitivity, low cytotoxicity, increased AM acetylmethyl ester can enter the cell well, after being sheared by the intracellular esterase stay in the cell to bind to calcium ions, produce strong fluorescence^[1].

Guide (The following is our recommended protocol. This protocol only provides guidance and should be modified according to your specific needs).
1. Preparation of Mag-Fluo-4 AM working solution
1.1 Preparation of stock solution
Dissolve Mag-Fluo-4 AM in anhydrous DMSO to prepare a 10 mM stock solution.
Note: It is recommended to aliquot the Mag-Fluo-4 AM stock solution and store at -20°C or -80°C, protected from light.
1.2 Preparation of working solution
Dilute the stock solution with pre-warmed HBSS to prepare a 1-10 μM Mag-Fluo-4 AM working solution.
Note: Please adjust the concentration of the Mag-Fluo-4 AM working solution according to actual conditions and prepare fresh before use.
2. Cell staining (suspension cells)
2.1 Collect cells by centrifugation, wash twice with PBS for 5 minutes each time. Adjust the cell density to 1×10⁶/mL
2.2 Add 1 mL Mag-Fluo-4 AM working solution and incubate at room temperature for 5-30 minutes.
2.3 Centrifuge at 400 g for 3-4 minutes and discard the supernatant.
2.4 Wash the cells twice with PBS for 5 minutes each time.
2.5 Resuspend the cells in 1 mL HBSS, then observe using a fluorescence microscope or flow cytometer.
3. Cell staining (adherent cells)
3.1 Culture adherent cells on sterile coverslips.
3.2 Remove the coverslips from the culture medium and aspirate excess medium.
3.3 Add 100 μL staining working solution, gently shake to ensure complete coverage of the cells, and incubate for 5-30 minutes.
3.4 Remove the staining working solution and wash with culture medium 2-3 times for 5 minutes each time, then observe using a fluorescence microscope.

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

817.65

C₃₇H₃₃F₂NO₁₈

1097709-31-3

Solid

Yellow to orange

515

475

O=C1C(F)=CC2=C(C3=CC=C(N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)C(OCC(OCOC(C)=O)=O)=C3)C4=C(C=C(OCOC(C)=O)C(F)=C4)OC2=C1

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

• [1]. Rossi AM, et al. Reliable measurement of free Ca2+ concentrations in the ER lumen using Mag-Fluo-4. Cell Calcium. 2020 May;87:102188.  [Content Brief]

  [2]. Gee KR, et al. Chemical and physiological characterization of fluo-4 Ca(2+)-indicator dyes. Cell Calcium. 2000 Feb;27(2):97-106.  [Content Brief]