Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>

Mag-Fluo-4 AM

Chemical structure for Mag-Fluo-4 AM
Chemical structure for Mag-Fluo-4 AM
Ordering information
Price ()
Catalog Number20401
Unit Size
Find Distributor
Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
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
Related products
Chemical Phosphorylation Reagent I (CPR I)
Cell Meter™ Mitochondrial Hydroxyl Radical Detection Kit *Red Fluorescence*
Cal Green™ 1, hexapotassium salt
Cal Green™ 1, AM [Equivalent to Calcium Green-1, AM]
Cal-590™-Dextran Conjugate *MW 3,000*
Cal-590™-Dextran Conjugate *MW 10,000*
Cal-590™ AM
Cal-590™, sodium salt
Cal-590™, potassium salt
Cal-630™ AM
Cal-630™, sodium salt
Cal-630™, potassium salt
Cal-630™-Dextran Conjugate *MW 3,000*
Cal-630™-Dextran Conjugate *MW 10,000*
Fluo-4 AM *Ultrapure Grade* *CAS 273221-67-3*
Fluo-4, Pentapotassium Salt
Cal Red™ R525/650 potassium salt
Cal Red™ R525/650 AM
Cal-520®-Dextran Conjugate *MW 3,000*
Cal-520®-Dextran Conjugate *MW 10,000*
Cal-520®-Biotin Conjugate
Cal-520®-Biocytin Conjugate
Cal-520® NHS Ester
Cal-520® maleimide
Fluo-3, AM *CAS 121714-22-5*
Fluo-3, AM *UltraPure grade* *CAS 121714-22-5*
Fluo-3, AM *Bulk package* *CAS 121714-22-5*
Fluo-3FF, AM *UltraPure grade* *Cell permeant*
Fluo-3, pentasodium salt
Fluo-3, pentapotassium salt
Fluo-3, pentaammonium salt
Fluo-3FF, pentapotassium salt
Fluo-8®, AM
Fluo-8®, sodium salt
Fluo-8®, potassium salt
Fluo-8H™, AM
Fluo-8H™, sodium salt
Fluo-8L™, AM
Fluo-8L™, sodium salt
Fluo-8L™, potassium salt
Fluo-8FF™, potassium salt
Fluo-8FF™, AM
Cal-520®, AM
Cal-520®, sodium salt
Cal-520®, potassium salt
Cal-520FF™, AM
Cal-520FF™, potassium salt
Screen Quest™ Fluo-8 Medium Removal Calcium Assay Kit *Optimized for Difficult Cell Lines*
Screen Quest™ Fluo-8 No Wash Calcium Assay Kit
Mag-Fluo-4 potassium salt
Fluo-2, potassium salt
Fluo-2, AM
Fluo-5F, AM *Cell permeant*
Fluo-5F, pentapotassium Salt *Cell impermeant*
Fluo-5N, AM *Cell permeant*
Fluo-5N, pentapotassium Salt *Cell impermeant*
Cal-520N™, AM
Cal-520N™, potassium salt
Screen Quest™ Fluo-4 No Wash Calcium Assay Kit
Calbryte™ 520 AM
Calbryte™ 520, potassium salt
Calbryte™ 590 AM
Calbryte™ 590, potassium salt
Calbryte™ 630 AM
Calbryte™ 630, potassium salt
Screen Quest™ Calbryte-520 Probenecid-Free and Wash-Free Calcium Assay Kit
Screen Quest™ Calbryte-590 Probenecid-Free and Wash-Free Calcium Assay Kit
Calbryte™-520L AM
Calbryte™-520L, potassium salt
Cal-500™, potassium salt
Cal-500™ AM
Cal-670™, potassium salt
Cal-670™-Dextran Conjugate *MW 3,000*
Cal-670™-Dextran Conjugate *MW 10,000*
Cal-770™, potassium salt
Cal-770™-Dextran Conjugate *MW 3,000*
Cal-770™-Dextran Conjugate *MW 10,000*
Calbryte™-520XL azide
Calbryte™-520XL, potassium salt
Calbryte™-520XL AM
Calbryte™-520XL-Dextran
Cal-520L®-Dextran Conjugate *MW 10,000*
RatioWorks™ Cal-520L®/Cy5-Dextran Conjugate *MW 10,000*
Cal-520L™ maleimide
RatioWorks™ Cal-520®/zFluor 647™ -Dextran Conjugate *MW 10,000*
Cal-590L® Dextran Conjugate *MW 10,000*
RatioWorks™ Cal-590L®/Cy5-Dextran Conjugate *MW 10,000*
Show More (87)

OverviewpdfSDSpdfProtocol


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
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
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. Prepare a dye working solution of 2 to 20 µM 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 to 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 solution, 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 into 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
/=x=

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

Citations


View all 7 citations: Citation Explorer
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
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
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
Hypocalcemia in sepsis: analysis of the subcellular distribution of Ca2+ in septic rats and LPS/TNF-$\alpha$-treated HUVECs
Authors: He, Wencheng and Huang, Lei and Luo, Hua and Zang, Yang and An, Youzhong and Zhang, Weixing
Journal: The Journal of Infection in Developing Countries (2020): 908--917
HuR regulates phospholamban expression in isoproterenol-induced cardiac remodelling
Authors: Hu, Han and Jiang, Mingyang and Cao, Yangpo and Zhang, Zhuojun and Jiang, Bin and Tian, Feng and Feng, Juan and Dou, Yali and Gorospe, Myriam and Zheng, Ming and others,
Journal: Cardiovascular research (2020): 944--955
HuR regulates phospholamban expression in isoproterenol-induced cardiac remodeling
Authors: Hu, Han and Jiang, Mingyang and Cao, Yangpo and Zhang, Zhuojun and Jiang, Bin and Tian, Feng and Feng, Juan and Dou, Yali and Gorospe, Myriam and Zheng, Ming and others, undefined
Journal: Cardiovascular research (2019)

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
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
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
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