Mag-520™ AM

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	579.46
Solvent	DMSO

Spectral properties

Excitation (nm)	506
Emission (nm)	525

Storage, safety and handling

Intended use	Research Use Only (RUO)
Storage	Freeze (< -15 °C); Minimize light exposure

Overview SDS Protocol

Platform

Flow cytometer

Excitation	488 nm laser
Emission	530/30 nm filter
Instrument specification(s)	FITC channel

Fluorescence microscope

Excitation	FITC filter set
Emission	FITC filter set
Recommended plate	Black wall/clear bottom

Example protocol

AT A GLANCE

Protocol summary

Grow cells as desired
Prepare and add Mag-520™ AM working solution to samples
Incubate samples at 37 °C for 15 to 45 minutes
Monitor the fluorescence intensity using flow cytometer with 530/30 nm filter (FITC channel) or using fluorescence microscopy with FITC filter set

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-520™ AM stock solution

Add appropriate amount of DMSO into Mag-520™ AM vial to make 2-5 mM Mag-520™ AM stock solution.
Note Mag-520™ AM working solution should be used promptly.

PREPARATION OF WORKING SOLUTION

Mag-520™ AM working solution

Prepare 5-50 µM Mag-520™ AM working solution in buffer of your choice.
Note Mag-520™ AM working solution should be used promptly.
Note The concentration of the Mag-520™ AM should be optimized for different cell types and conditions.

SAMPLE EXPERIMENTAL PROTOCOL

The following protocol can be used as a guideline and should be optimized according to the needs.

Grow cells as desired.
Remove the treatment and wash the cells with buffer of your choice such as DPBS.
Add Mag-520™ AM working solution and incubate the samples for 15-45 minutes at 37 °C incubator.
Note Optimal time for incubation needs to be determined carefully.
Remove the working solution and wash cells with buffer of your choice.
Resuspend cells in buffer of your choice.
Add stimulant to stimulate the cells and monitor the fluorescence intensity with flow cytometer using 530/30 nm filter (FITC channel) or fluorescence microscope with FITC filter set.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Mag-520™ AM to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	172.574 µL	862.872 µL	1.726 mL	8.629 mL	17.257 mL
5 mM	34.515 µL	172.574 µL	345.149 µL	1.726 mL	3.451 mL
10 mM	17.257 µL	86.287 µL	172.574 µL	862.872 µL	1.726 mL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
	/		=		x		=

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Excitation (nm)	506
Emission (nm)	525

Product Family

Name	Excitation (nm)	Emission (nm)	Quantum yield
Cal-520®, AM	492	515	0.75¹
Calbryte™ 520 AM	493	515	0.75¹
SoNa™ 520 AM	491	511	-

Images

Figure 1. In vitro detection of Mg⁺². HL-60 cells were stained with AM form of Mag-520™ (5 uM) and mag-fluo-4 (5 uM) for 30 minutes and stimulated with FCCP (20 uM) for 30 minutes and response was recorded for end point analysis. Response was recorded using NovoCyte™ 3000 Flow Cytometer with 530/30 nm filter and 488 nm laser.

Figure 2. The FCCP induced intracellular Magnesium release was measured by Mag-520^TM. Cells were incubated with Mag-520™ AM dye for 30 min at 37 °C before 20 µM FCCP was added into the cells. The baseline was acquired and the rest of the cells were analyzed after the addition of FCCP. The response was measured over time. The analysis was done on NovoCyte™ 3000 Flow Cytometer.

Citations

View all 15 citations: Citation Explorer

A Highly Selective Green Fluorescent Magnesium Indicator for Intracellular Magnesium Ion Analysis
Authors: Patel, Deven and Zhao, Qin and Guo, Haitao and Peng, Ruogu and Liu, Jixiang and Liao, Jinfang and Diwu, Zhenjun
Journal: Biophysical Journal (2020): 273a

A highly selective fluorescent probe for the intracellular measurement of magnesium ion
Authors: Patel, Deven and Peng, Ruogu and Guo, Haitao and Lin, Raechel and Zhao, Qin and Liao, Jinfang and Diwu, Zhenjun
Journal: Analytical Biochemistry (2020): 113910

Fluorescence lifetime imaging of intracellular magnesium content in live cells
Authors: Sargenti, A., C and eo, A., Farruggia, G., D'Andrea, C., Cappadone, C., Malucelli, E., Valentini, G., Taroni, P., Iotti, S.
Journal: Analyst (2019): 1876-1880

Synthesis of a highly Mg(2+)-selective fluorescent probe and its application to quantifying and imaging total intracellular magnesium
Authors: Sargenti, A., Farruggia, G., Zaccheroni, N., Marraccini, C., Sgarzi, M., Cappadone, C., Malucelli, E., Procopio, A., Prodi, L., Lombardo, M., Iotti, S.
Journal: Nat Protoc (2017): 461-471

A novel fluorescent chemosensor allows the assessment of intracellular total magnesium in small samples
Authors: Sargenti, A., Farruggia, G., Malucelli, E., Cappadone, C., Merolle, L., Marraccini, C., Andreani, G., Prodi, L., Zaccheroni, N., Sgarzi, M., Trombini, C., Lombardo, M., Iotti, S.
Journal: Analyst (2014): 1201-7

Intracellular magnesium detection by fluorescent indicators
Authors: Trapani, V., Schweigel-Rontgen, M., Cittadini, A., Wolf, F. I.
Journal: Methods Enzymol (2012): 421-44

Development of fluorescent probes for intracellular magnesium measurement and their applications to analysis of cellular mechanism
Authors: Shindo, Y., Oka, K., Komatsu, H., Suzuki, K.
Journal: Tanpakushitsu Kakusan Koso (2007): 1552-7

Mitochondria are intracellular magnesium stores: investigation by simultaneous fluorescent imagings in PC12 cells
Authors: Kubota, T., Shindo, Y., Tokuno, K., Komatsu, H., Ogawa, H., Kudo, S., Kitamura, Y., Suzuki, K., Oka, K.
Journal: Biochim Biophys Acta (2005): 19-28

Investigation of intracellular magnesium mobilization pathways I PC12 cells B simultaneous Mg-Ca fluorescent imaging
Authors: Kubota, T., Shindo, Y., Tokuno, K., Komatsu, H., Ogawa, H., Kitamura, Y., Suzuki, K., Oka, K.
Journal: J Am Coll Nutr (2004): 742S-4S

Design and synthesis of highly sensitive and selective fluorescein-derived magnesium fluorescent probes and application to intracellular 3D Mg2+ imaging
Authors: Komatsu, H., Iwasawa, N., Citterio, D., Suzuki, Y., Kubota, T., Tokuno, K., Kitamura, Y., Oka, K., Suzuki, K.
Journal: J Am Chem Soc (2004): 16353-60