Mag-Fluo-4 AM
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Dissociation constant (Kd, nM) | 22000 |
Molecular weight | 817.65 |
Solvent | DMSO |
Extinction coefficient (cm -1 M -1) | 82000 |
Excitation (nm) | 495 |
Emission (nm) | 528 |
Quantum yield | 0.161 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
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 |
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 |
Fluorescence microplate reader
Excitation | 490 |
Emission | 525 |
Cutoff | 515 |
Recommended plate | Black 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
Prepare a 2 to 5 mM stock solution of Mag-Fluo-4 AM in high-quality, anhydrous DMSO.
PREPARATION OF 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 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.
- Prepare cells in growth medium overnight.
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.
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.
- 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.
- 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
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 122.302 µL | 611.509 µL | 1.223 mL | 6.115 mL | 12.23 mL |
5 mM | 24.46 µL | 122.302 µL | 244.603 µL | 1.223 mL | 2.446 mL |
10 mM | 12.23 µL | 61.151 µL | 122.302 µL | 611.509 µL | 1.223 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum

Spectral properties
Extinction coefficient (cm -1 M -1) | 82000 |
Excitation (nm) | 495 |
Emission (nm) | 528 |
Quantum yield | 0.161 |
Citations
Authors: Chang, Jiayin and Yan, Shihai and Geng, Zhirong and Wang, Zhilin
Journal: Toxicology and Applied Pharmacology (2023): 116511
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
Authors: Plasterer, Cody and Semenikhina, Marharyta and Tsaih, Shirng-Wern and Flister, Michael J and Palygin, Oleg
Journal: Molecular Medicine (2023): 1--12
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
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
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
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
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
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
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
References
Authors: Fern, undefined and ez-Tenorio M, Niggli E.
Journal: Cell Calcium. (2016)
Authors: Wang H, Wang ZK, Jiao P, Zhou XP, Yang DB, Wang ZY, Wang L.
Journal: Toxicology (2015): 137
Authors: Ainbinder A, Boncompagni S, Protasi F, Dirksen RT.
Journal: Cell Calcium (2015): 14
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Journal: Toxicol Lett (2014): 52
Authors: Calderon JC, Bolanos P, Caputo C.
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Authors: Trapani V, Arduini D, Luongo F, Wolf FI.
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Authors: Tovey SC, Taylor CW.
Journal: Cold Spring Harb Protoc (2013): 930
Authors: Kojima A, Kitagawa H, Omatsu-Kanbe M, Matsuura H, Nosaka S.
Journal: Br J Anaesth (2012): 352
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Journal: J Physiol (2012): 575
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