Fura-2, AM *UltraPure Grade* *CAS 108964-32-5*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
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Quotation | Request |
International | See distributors |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Dissociation constant (Kd, nM) | 145 |
Molecular weight | 1001.86 |
Solvent | DMSO |
Spectral properties
Excitation (nm) | 336 |
Emission (nm) | 505 |
Storage, safety and handling
Certificate of Origin | Download PDF |
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 |
Alternative formats
Fura-2, AM *CAS 108964-32-5* |
Overview | ![]() ![]() |
See also: Calcium Indicators, Intracellular Ions, Physiological Probes, Ratiometric Calcium Indicators
CAS 108964-32-5 | Molecular weight 1001.86 | Dissociation constant (Kd, nM) 145 | Excitation (nm) 336 | Emission (nm) 505 |
Among the ratiometric calcium indicators, Fura-2 and Indo-1 are most commonly used. Fura-2 is excitation-ratioable while Indo-1 is emission-ratioable. Fura-2 is preferred for ratio-imaging microscopy, in which it is more practical to change excitation wavelengths than emission wavelengths. Upon binding Ca2+, Fura-2 exhibits an absorption shift that can be observed by scanning the excitation spectrum between 300 and 400 nm, while monitoring the emission at ~510 nm. Fura-2, AM is a cell-permeable calcium indicator that is emission-ratiometric and UV light'excitable. This AM ester form can be loaded into live cells noninvasively.
Platform
Fluorescence microscope
Excitation | Fura 2 filter set |
Emission | Fura 2 filter set |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 340, 380 |
Emission | 510 |
Cutoff | 475 |
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.
Fura-2 AM *UltraPure Grade* Stock Solution
Prepare a 2 to 5 mM stock solution of Fura-2 AM in high-quality, anhydrous DMSO.PREPARATION OF WORKING SOLUTION
Fura-2 AM *UltraPure Grade* Working Solution
On the day of the experiment, either dissolve Fura-2 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, 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 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 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.
- Prepare cells in growth medium overnight.
- On the next day, add 1X Fura-2 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. - 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 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 Fura-2, AM *UltraPure Grade* *CAS 108964-32-5* 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 | 99.814 µL | 499.072 µL | 998.143 µL | 4.991 mL | 9.981 mL |
5 mM | 19.963 µL | 99.814 µL | 199.629 µL | 998.143 µL | 1.996 mL |
10 mM | 9.981 µL | 49.907 µL | 99.814 µL | 499.072 µL | 998.143 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Product Family
Name | Excitation (nm) | Emission (nm) |
Fura-8FF™, AM | 354 | 524 |
Fura-FF, AM [Fura-2FF, AM] *CAS 348079-12-9* | 336 | 505 |
Fura Red, AM *CAS 149732-62-7* | 435 | 639 |
Fura-8™, AM | 354 | 524 |
Rhod-2, AM *CAS#: 145037-81-6* | 553 | 577 |
Rhod-2, AM *UltraPure Grade* *CAS#: 145037-81-6* | 553 | 577 |
Fura-10™, AM | 354 | 524 |
Images
Citations
View all 13 citations: Citation Explorer
Inhibitory responses to retinohypothalamic tract stimulation in the circadian clock of the diurnal rodent Rhabdomys pumilio
Authors: Schoonderwoerd, Robin A and de Torres Guti{\'e}rrez, Pablo and Blommers, Ruben and van Beurden, Anouk W and Coenen, Tineke CJJ and Klett, Nathan J and Michel, Stephan H and Meijer, Johanna H
Journal: The FASEB Journal (2022): e22415
Authors: Schoonderwoerd, Robin A and de Torres Guti{\'e}rrez, Pablo and Blommers, Ruben and van Beurden, Anouk W and Coenen, Tineke CJJ and Klett, Nathan J and Michel, Stephan H and Meijer, Johanna H
Journal: The FASEB Journal (2022): e22415
IKCa channels control breast cancer metabolism including AMPK-driven autophagy
Authors: Gross, Dominic and Bischof, Helmut and Maier, Selina and Sporbeck, Katharina and Birkenfeld, Andreas L and Malli, Roland and Ruth, Peter and Proikas-Cezanne, Tassula and Lukowski, Robert
Journal: Cell death \& disease (2022): 1--14
Authors: Gross, Dominic and Bischof, Helmut and Maier, Selina and Sporbeck, Katharina and Birkenfeld, Andreas L and Malli, Roland and Ruth, Peter and Proikas-Cezanne, Tassula and Lukowski, Robert
Journal: Cell death \& disease (2022): 1--14
Transmembrane Domain 3 Is a Transplantable Pharmacophore in the Photodynamic Activation of Cholecystokinin 1 Receptor
Authors: Li, Yuan and Cui, Zong Jie
Journal: ACS Pharmacology \& Translational Science (2022)
Authors: Li, Yuan and Cui, Zong Jie
Journal: ACS Pharmacology \& Translational Science (2022)
Catalpol attenuates renal injury by regulating oxidative stress and inflammation response
Authors: Liu, Zhihui and Wang, Yu and Zhou, Chong and Xu, Qingyang and Gao, Hongxin and Huo, Mohan and Jiang, Xiaowen and Yu, Wenhui
Journal: (2022)
Authors: Liu, Zhihui and Wang, Yu and Zhou, Chong and Xu, Qingyang and Gao, Hongxin and Huo, Mohan and Jiang, Xiaowen and Yu, Wenhui
Journal: (2022)
NanoLuc Bioluminescence-Driven Photodynamic Activation of Cholecystokinin 1 Receptor with Genetically-Encoded Protein Photosensitizer MiniSOG
Authors: Li, Yuan and Cui, Zong Jie
Journal: International Journal of Molecular Sciences (2020): 3763
Authors: Li, Yuan and Cui, Zong Jie
Journal: International Journal of Molecular Sciences (2020): 3763
Photodynamic Activation of Cholecystokinin 1 Receptor with Different Genetically Encoded Protein Photosensitizers and from Varied Subcellular Sites
Authors: Li, Yuan and Cui, Zong Jie
Journal: Biomolecules (2020): 1423
Authors: Li, Yuan and Cui, Zong Jie
Journal: Biomolecules (2020): 1423
Aspirin eugenol ester attenuates oxidative injury of vascular endothelial cells by regulating NOS and Nrf2 signaling pathways
Authors: Huang, Mei-Zhou and Yang, Ya-Jun and Liu, Xi-Wang and Qin, Zhe and Li, Jian-Yong
Journal: British Journal of Pharmacology (2019)
Authors: Huang, Mei-Zhou and Yang, Ya-Jun and Liu, Xi-Wang and Qin, Zhe and Li, Jian-Yong
Journal: British Journal of Pharmacology (2019)
Effect of PEGylated Magnetic PLGA-PEI Nanoparticles on Primary Hippocampal Neurons: Reduced Nano-neurotoxicity and Enhanced Transfection Efficiency with Magnetofection
Authors: Cui, Yanna and Li, Xiao and Zeljic, Kristina and Shan, Shifang and Qiu, Zilong and Wang, Zheng
Journal: ACS Applied Materials & Interfaces (2019)
Authors: Cui, Yanna and Li, Xiao and Zeljic, Kristina and Shan, Shifang and Qiu, Zilong and Wang, Zheng
Journal: ACS Applied Materials & Interfaces (2019)
Periprosthetic hypoxia as consequence of TRPM7 mediated cobalt influx in osteoblasts
Authors: Römmelt, Constantin and Munsch, Thomas and Drynda, Andreas and Lessmann, Volkmar and Lohmann, Christoph H and Bertr, undefined and , Jessica
Journal: Journal of Biomedical Materials Research Part B: Applied Biomaterials (2018)
Authors: Römmelt, Constantin and Munsch, Thomas and Drynda, Andreas and Lessmann, Volkmar and Lohmann, Christoph H and Bertr, undefined and , Jessica
Journal: Journal of Biomedical Materials Research Part B: Applied Biomaterials (2018)
An essential role of NAD (P) H oxidase 2 in UVA-induced calcium oscillations in mast cells
Authors: Li, Zhi Ying and Jiang, Wen Yi and Cui, Zong Jie
Journal: Photochemical \& Photobiological Sciences (2015): 414--428
Authors: Li, Zhi Ying and Jiang, Wen Yi and Cui, Zong Jie
Journal: Photochemical \& Photobiological Sciences (2015): 414--428
References
View all 119 references: Citation Explorer
Load of calcium probe Fura -2/AM in Escherichia coli cells
Authors: Shao M, Wang HM, Liu ZH, Shen P, Cai RX.
Journal: Wei Sheng Wu Xue Bao (2005): 805
Authors: Shao M, Wang HM, Liu ZH, Shen P, Cai RX.
Journal: Wei Sheng Wu Xue Bao (2005): 805
An Excel-based model of Ca2+ diffusion and fura 2 measurements in a spherical cell
Authors: McHugh JM, Kenyon JL.
Journal: Am J Physiol Cell Physiol (2004): C342
Authors: McHugh JM, Kenyon JL.
Journal: Am J Physiol Cell Physiol (2004): C342
Problems caused by high concentration of ATP on activation of the P2X7 receptor in bone marrow cells loaded with the Ca2+ fluorophore fura-2
Authors: Paredes-Gamero EJ, Franca JP, Moraes AA, Aguilar MO, Oshiro ME, Ferreira AT.
Journal: J Fluoresc (2004): 711
Authors: Paredes-Gamero EJ, Franca JP, Moraes AA, Aguilar MO, Oshiro ME, Ferreira AT.
Journal: J Fluoresc (2004): 711
Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2
Authors: McConnell G, Riis E.
Journal: Phys Med Biol (2004): 4757
Authors: McConnell G, Riis E.
Journal: Phys Med Biol (2004): 4757
Abnormal spectra alteration observed in Triton calibration method for measuring [Ca2+]i with fluorescence indicator, fura-2
Authors: Xu T, Yang W, Huo XL, Song T.
Journal: J Biochem Biophys Methods (2004): 219
Authors: Xu T, Yang W, Huo XL, Song T.
Journal: J Biochem Biophys Methods (2004): 219
Two-photon microscopy of fura-2-loaded cardiac myocytes with an all-solid-state tunable and visible femtosecond laser source
Authors: McConnell G, Smith GL, Girkin JM, Gurney AM, Ferguson AI.
Journal: Opt Lett (2003): 1742
Authors: McConnell G, Smith GL, Girkin JM, Gurney AM, Ferguson AI.
Journal: Opt Lett (2003): 1742
AMPA-induced Ca(2+) influx in cultured rat cortical nonpyramidal neurones: pharmacological characterization using fura-2 microfluorimetry
Authors: Fischer W, Franke H, Scheibler P, Allgaier C, Illes P.
Journal: Eur J Pharmacol (2002): 53
Authors: Fischer W, Franke H, Scheibler P, Allgaier C, Illes P.
Journal: Eur J Pharmacol (2002): 53
Selective measurement of endothelial or smooth muscle [Ca(2+)](i) in pressurized/perfused cerebral arteries with fura-2
Authors: Marrelli SP., undefined
Journal: J Neurosci Methods (2000): 145
Authors: Marrelli SP., undefined
Journal: J Neurosci Methods (2000): 145
Excitation wavelengths for fura 2 provide a linear relationship between [Ca(2+)] and fluorescence ratio
Authors: Palmer BM, Moore RL.
Journal: Am J Physiol Cell Physiol (2000): C1278
Authors: Palmer BM, Moore RL.
Journal: Am J Physiol Cell Physiol (2000): C1278
Tyrosine kinase inhibitors and Ca2+ signaling: direct interactions with fura-2
Authors: Berts A, Minneman KP.
Journal: Eur J Pharmacol (2000): 35
Authors: Berts A, Minneman KP.
Journal: Eur J Pharmacol (2000): 35
Application notes
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A Meta-Analysis of Common Calcium Indicators
A New Red Fluorescent & Robust Screen Quest™ Rhod-4™ Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
A Meta-Analysis of Common Calcium Indicators
A New Red Fluorescent & Robust Screen Quest™ Rhod-4™ Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets