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Mag-Fluo-4 AM

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Physical properties
Dissociation constant (Kd, nM)22000
Molecular weight817.65
SolventDMSO
Spectral properties
Extinction coefficient (cm -1 M -1)82000
Excitation (nm)495
Emission (nm)528
Quantum yield0.161
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
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Show More (82)

OverviewpdfSDSpdfProtocol


See also: Fluoresceins
Molecular weight
817.65
Dissociation constant (Kd, nM)
22000
Extinction coefficient (cm -1 M -1)
82000
Excitation (nm)
495
Emission (nm)
528
Quantum yield
0.161
The cell-permeant Mag-Fluo-4 AM is an analog of Fluo-4 AM with a Kd for Mg ion of 4.7 mM and a Kd for Ca ion of 22 µM, making it useful as an intracellular Mg ion indicator as well as a low-affinity Ca ion indicator. This low-affinity fluorescent Ca ion indicator has been used to accurately track the kinetics of the spatially averaged free Ca ion transient in skeletal muscle. Mag-fluo-4 yields reliable kinetic information about the spatially averaged free Ca ion transient in skeletal muscle.

Platform


Flow cytometer

Excitation488 nm laser
Emission530/30 nm filter
Instrument specification(s)FITC channel

Fluorescence microscope

ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall/clear bottom

Fluorescence microplate reader

Excitation490
Emission525
Cutoff515
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-Fluo-4 AM Stock Solution
  1. Prepare a 2 to 5 mM stock solution of Mag-Fluo-4 AM in high-quality, anhydrous DMSO.

PREPARATION OF WORKING SOLUTION

Mag-Fluo-4 AM Working Solution
  1. On the day of the experiment, either dissolve Mag-Fluo-4 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.

  2. Prepare a 2 to 20 µM Mag-Fluo-4 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-Fluo-4 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-Fluo-4 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-Fluo-4 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 FITC filter set or a fluorescence plate reader containing a programmable liquid handling system such as an FDSS, FLIPR, or FlexStation, at Ex/Em = 490/525 nm cutoff 515 nm.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Mag-Fluo-4 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 mg0.5 mg1 mg5 mg10 mg
1 mM122.302 µL611.509 µL1.223 mL6.115 mL12.23 mL
5 mM24.46 µL122.302 µL244.603 µL1.223 mL2.446 mL
10 mM12.23 µL61.151 µL122.302 µL611.509 µL1.223 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


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Extinction coefficient (cm -1 M -1)82000
Excitation (nm)495
Emission (nm)528
Quantum yield0.161

Images


Citations


View all 13 citations: Citation Explorer
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Journal: Pharmaceuticals (2024): 805
Mitochondrial-dependent oxidative phosphorylation is key for postnatal metabolic adaptation of alveolar macrophages in the lung
Authors: Zhang, Jun and Peng, Yu and Song, Haosen and Liu, Siqi and Li, Chuanwei and Zhang, Yi and Shi, Xiaowei and Guo, Huifang and Xu, Yingping
Journal: International Immunopharmacology (2024): 112012
NNAT is a novel mediator of oxidative stress that suppresses ER+ breast cancer
Authors: Plasterer, Cody and Semenikhina, Marharyta and Tsaih, Shirng-Wern and Flister, Michael J and Palygin, Oleg
Journal: Molecular Medicine (2023): 1--12
Berbamine promotes macrophage autophagy to clear Mycobacterium tuberculosis by regulating the ROS/Ca2+ axis
Authors: Zhang, Su and Zhou, Xuefeng and Ou, Min and Fu, Xiangdong and Lin, Qiao and Tao, Xiaoyu and Wang, Zhaoqin and Liu, Aimei and Li, Guobao and Xu, Yuzhong and others,
Journal: mBio (2023): e00272--23
Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant
Authors: Qiu, Yue and Wang, Hongyang and Fan, Mingjie and Pan, Huaye and Guan, Jing and Jiang, Yangwei and Jia, Zexiao and Wu, Kaiwen and Zhou, Hui and Zhuang, Qianqian and others,
Journal: Cell Death \& Disease (2023): 375
The interaction between Hsp90-mediated unfolded protein response and autophagy contributes to As3+/Se4+ combination-induced apoptosis of acute promyelocytic leukemia cells
Authors: Chang, Jiayin and Yan, Shihai and Geng, Zhirong and Wang, Zhilin
Journal: Toxicology and Applied Pharmacology (2023): 116511
SYNJ2BP Improves the Production of Lentiviral Envelope Protein by Facilitating the Formation of Mitochondrion-Associated Endoplasmic Reticulum Membrane
Authors: Duan, Yingyi and Wang, Xinhui and Sun, Kehui and Lin, Yuezhi and Wang, Xuefeng and Chen, Kewei and Yang, Guangpu and Wang, Xiaojun and Du, Cheng
Journal: Journal of Virology (2022): e00549--22
Reticulon 2 promotes gastric cancer metastasis via activating endoplasmic reticulum Ca2+ efflux-mediated ERK signalling
Authors: Song, Shushu and Liu, Bo and Zeng, Xiaoqing and Wu, Yingying and Chen, Hao and Wu, Hao and Gu, Jianxin and Gao, Xiaodong and Ruan, Yuanyuan and Wang, Hongshan
Journal: Cell death \& disease (2022): 1--13
Mitochondrial Calcium-Triggered Oxidative Stress and Developmental Defects in Dopaminergic Neurons Differentiated from Deciduous Teeth-Derived Dental Pulp Stem Cells with MFF Insufficiency
Authors: Sun, Xiao and Dong, Shuangshan and Kato, Hiroki and Kong, Jun and Ito, Yosuke and Hirofuji, Yuta and Sato, Hiroshi and Kato, Takahiro A and Sakai, Yasunari and Ohga, Shouichi and others,
Journal: Antioxidants (2022): 1361
The three-spot seahorse-derived peptide PAGPRGPA attenuates ethanol-induced oxidative stress in LO2 cells through MAPKs, the Keap1/Nrf2 signalling pathway and amino acid metabolism
Authors: Shi, Jie and Zhou, Xin and Zhao, Ying and Tang, Xuemei and Feng, Lu and Wang, Boyuan and Chen, Jian
Journal: Food \& Function (2021): 1672--1687

References


View all 25 references: Citation Explorer
Real-time intra-store confocal Ca2+ imaging in isolated mouse cardiomyocytes
Authors: Fern, undefined and ez-Tenorio M, Niggli E.
Journal: Cell Calcium. (2016)
Redistribution of subcellular calcium and its effect on apoptosis in primary cultures of rat proximal tubular cells exposed to lead
Authors: Wang H, Wang ZK, Jiao P, Zhou XP, Yang DB, Wang ZY, Wang L.
Journal: Toxicology (2015): 137
Role of Mitofusin-2 in mitochondrial localization and calcium uptake in skeletal muscle
Authors: Ainbinder A, Boncompagni S, Protasi F, Dirksen RT.
Journal: Cell Calcium (2015): 14
EGF stimulates Mg(2+) influx in mammary epithelial cells
Authors: Trapani V, Arduini D, Luongo F, Wolf FI.
Journal: Biochem Biophys Res Commun (2014): 572
TRPC1 is involved in Ca(2)(+) influx and cytotoxicity following Pb(2)(+) exposure in human embryonic kidney cells
Authors: Zhang H, Li W, Xue Y, Zou F.
Journal: Toxicol Lett (2014): 52
Tetanic Ca2+ transient differences between slow- and fast-twitch mouse skeletal muscle fibres: a comprehensive experimental approach
Authors: Calderon JC, Bolanos P, Caputo C.
Journal: J Muscle Res Cell Motil (2014): 279
High-throughput functional assays of IP3-evoked Ca2+ release
Authors: Tovey SC, Taylor CW.
Journal: Cold Spring Harb Protoc (2013): 930
Presence of store-operated Ca2+ entry in C57BL/6J mouse ventricular myocytes and its suppression by sevoflurane
Authors: Kojima A, Kitagawa H, Omatsu-Kanbe M, Matsuura H, Nosaka S.
Journal: Br J Anaesth (2012): 352
Measurement and simulation of myoplasmic calcium transients in mouse slow-twitch muscle fibres
Authors: Hollingworth S, Kim MM, Baylor SM.
Journal: J Physiol (2012): 575
TRPC1, STIM1, and ORAI influence signal-regulated intracellular and endoplasmic reticulum calcium dynamics in human myometrial cells
Authors: Murtazina DA, Chung D, Ulloa A, Bryan E, Galan HL, Sanborn BM.
Journal: Biol Reprod (2011): 315