Mag-indo-1/AM

Mag-indo-1/AM is a cell-permeable fluorescent indicator for Mg2+ and also for Ca2+ (Ex=340-390 nm, Em=410-490 nm). Mag-indo-1/AM can be used to determine intracellular 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). Measurement of intracellular Ca2+[1]: 1. Incubate 3 mL cell suspension with 10 µM Mag-indo-1/AM for 15 min at 30℃. 2. Centrifuge the cells for 1 min at 500 rpm and resuspend in 5 mL isolation buffer (containing 1 mM CaCl2). 3. Add 1 mM probenecid to the cell suspension to prevent dye leakage from the cytosol and store the cells (Mag-indo-1/AM-loaded cells) at room temperature until use. 4. Incubate the Mag-indo-1/AM-loaded cells at room temperature for 2 h, shake gently at 37℃ for 1.5 h, centrifuge the pellet, resuspend and store at room temperature. 6. Place a small portion of the loaded cells in an experimental chamber which is mounted on the stage of an inverted microscope. 7. Measure fluorescence of Mag-indo-1/AM (Ex=340-390 nm, Em=410 and 490 nm (the peak emissions of the Ca2+ bound and Ca2+ free forms of the indicator)). Fluorescence of the whole cell is collected on line by a photon counting system.

[1]. Griffiths EJ. Calcium handling and cell contraction in rat cardiomyocytes depleted of intracellular magnesium. Cardiovasc Res. 2000 Jul;47(1):116-23.

[2]. Morelle B, et al. Measurement of intracellular magnesium concentration in 3T3 fibroblasts with the fluorescent indicator Mag-indo-1. Anal Biochem. 1994 Apr;218(1):170-6.