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Mag-Fura-2, AM *Cell-permeant*

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Physical properties
Molecular weight722.56
SolventDMSO
Spectral properties
Excitation (nm)336
Emission (nm)505
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


CAS
130100-20-8
Molecular weight
722.56
Excitation (nm)
336
Emission (nm)
505
Mag-Fura-2, AM is an intracellular magnesium indicator that is ratiometric and UV light-excitable. It has the spectral properties that closely match Fura-2. This acetoxymethyl (AM) ester form is useful for noninvasive intracellular loading. It is also used for measuring high level of calcium ion in live cells.

Platform


Fluorescence microscope

ExcitationFura 2 filter set
EmissionFura 2 filter set
Recommended plateBlack wall/clear bottom

Fluorescence microplate reader

Excitation340, 380
Emission510
Cutoff475
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling

Example protocol


PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

Mag-Fura-2 AM Stock Solution
  1. Prepare a 2 to 5 mM stock solution of Mag-Fura-2 AM in high-quality, anhydrous DMSO.

PREPARATION OF WORKING SOLUTION

Mag-Fura-2 AM Working Solution
  1. On the day of the experiment, either dissolve Mag-Fura-2 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.

  2. Prepare a 2 to 20 µM Mag-Fura-2 AM working solution in a buffer of your choice (e.g., Hanks and Hepes buffer) with 0.04% Pluronic® F-127. For most cell lines, Mag-Fura-2 AM at a final concentration of 4-5 μM is recommended. The exact concentration of indicators required for cell loading must be determined empirically.

    Note: The nonionic detergent Pluronic® F-127 is sometimes used to increase the aqueous solubility of Mag-Fura-2 AM. A variety of Pluronic® F-127 solutions can be purchased from AAT Bioquest.

    Note: If your cells contain organic anion-transporters, probenecid (1-2 mM) may be added to the dye working solution (final in well concentration will be 0.5-1 mM) to reduce leakage of the de-esterified indicators. A variety of ReadiUse™ Probenecid products, including water-soluble, sodium salt, and stabilized solutions, can be purchased from AAT Bioquest.

SAMPLE EXPERIMENTAL PROTOCOL

Following is our recommended protocol for loading AM esters into live cells. This protocol only provides a guideline and should be modified according to your specific needs.

  1. Prepare cells in growth medium overnight.
  2. On the next day, add 1X Mag-Fura-2 AM working solution to your cell plate.

    Note: If your compound(s) interfere with the serum, replace the growth medium with fresh HHBS buffer before dye-loading.

  3. Incubate the dye-loaded plate in a cell incubator at 37 °C for 30 to 60 minutes.

    Note: Incubating the dye for longer than 1 hour can improve signal intensities in certain cell lines.

  4. Replace the dye working solution with HHBS or buffer of your choice (containing an anion transporter inhibitor, such as 1 mM probenecid, if applicable) to remove any excess probes.
  5. Add the stimulant as desired and simultaneously measure fluorescence using either a fluorescence microscope equipped with a Fura 2 filter set or a fluorescence plate reader containing a programmable liquid handling system such as a FlexStation, at Ex/Em1 = 340/510 nm cutoff 475 nm and Ex/Em2 = 380/510 nm cutoff 475 nm.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Mag-Fura-2, AM *Cell-permeant* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM138.397 µL691.984 µL1.384 mL6.92 mL13.84 mL
5 mM27.679 µL138.397 µL276.794 µL1.384 mL2.768 mL
10 mM13.84 µL69.198 µL138.397 µL691.984 µL1.384 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum


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spectrum

Spectral properties

Excitation (nm)336
Emission (nm)505

Images


References


View all 15 references: Citation Explorer
Tuning the Spectroscopic Properties of Ratiometric Fluorescent Metal Indicators: Experimental and Computational Studies on Mag-Fura-2 and Analogues
Authors: Zhang G, Jacquemin D, Buccella D.
Journal: J Phys Chem B. (2017)
Fluorescence measurements of free [Mg2+] by use of mag-fura 2 in Salmonella enterica
Authors: Froschauer EM, Kolisek M, Dieterich F, Schweigel M, Schweyen RJ.
Journal: FEMS Microbiol Lett (2004): 49
Ionic selectivity of low-affinity ratiometric calcium indicators: mag-Fura-2, Fura-2FF and BTC
Authors: Hyrc KL, Bownik JM, Goldberg MP.
Journal: Cell Calcium (2000): 75
Characterization of Mg2+ transport in brush border membrane vesicles of rabbit ileum studied with mag-fura-2
Authors: Juttner R, Ebel H.
Journal: Biochim Biophys Acta (1998): 51
Intracellular Mg2+ regulation in voltage-clamped Helix aspersa neurones measured with mag-fura-2 and Mg(2+)-sensitive microelectrodes
Authors: Kennedy HJ., undefined
Journal: Exp Physiol (1998): 449
Mag-Fura-2 (Furaptra) exhibits both low (microM) and high (nM) affinity for Ca2+
Authors: Martinez-Zaguilan R, Parnami J, Martinez GM.
Journal: Cell Physiol Biochem (1998): 158
Monitoring of Ca2+ release from intracellular stores in permeabilized rat parotid acinar cells using the fluorescent indicators Mag-fura-2 and calcium green C18
Authors: Tojyo Y, Tanimura A, Matsumoto Y.
Journal: Biochem Biophys Res Commun (1997): 189
Intrasynaptosomal free Mg2+ concentration measured with the fluorescent indicator mag-fura-2: modulation by Na+ gradient and by extrasynaptosomal ATP
Authors: Garcia-Martin E, Martin-Romero FJ, Gutierrez-Merino C.
Journal: J Neurochem (1995): 2757
Effects of monovalent cations, pH and temperature on the dissociation constant (KD) for the fluorescent indicator mag-fura-2 at different excitation wavelengths
Authors: Howarth FC, Singh J, Waring JJ, Hustler BI, Bailey M.
Journal: Magnes Res (1995): 299
Interactions of polyamines in the measurement of free magnesium concentration by mag-fura-2 and 31P-NMR
Authors: Gunther T, Vormann J, Konstanczak P, Schafer A.
Journal: Biochim Biophys Acta (1994): 281