Mag-Fura-2, AM *Cell-permeant*
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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 |
Molecular weight | 722.56 |
Solvent | DMSO |
Excitation (nm) | 336 |
Emission (nm) | 505 |
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 | SDSProtocol |
CAS 130100-20-8 | Molecular weight 722.56 | Excitation (nm) 336 | Emission (nm) 505 |
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
Prepare a 2 to 5 mM stock solution of Mag-Fura-2 AM in high-quality, anhydrous DMSO.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Mag-Fura-2 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Mag-Fura-2 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-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 Mag-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 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-Fura-2 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 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
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 138.397 µL | 691.984 µL | 1.384 mL | 6.92 mL | 13.84 mL |
5 mM | 27.679 µL | 138.397 µL | 276.794 µL | 1.384 mL | 2.768 mL |
10 mM | 13.84 µL | 69.198 µL | 138.397 µL | 691.984 µL | 1.384 mL |
Molarity calculator
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References
Authors: Zhang G, Jacquemin D, Buccella D.
Journal: J Phys Chem B. (2017)
Authors: Froschauer EM, Kolisek M, Dieterich F, Schweigel M, Schweyen RJ.
Journal: FEMS Microbiol Lett (2004): 49
Authors: Hyrc KL, Bownik JM, Goldberg MP.
Journal: Cell Calcium (2000): 75
Authors: Juttner R, Ebel H.
Journal: Biochim Biophys Acta (1998): 51
Authors: Kennedy HJ., undefined
Journal: Exp Physiol (1998): 449
Authors: Martinez-Zaguilan R, Parnami J, Martinez GM.
Journal: Cell Physiol Biochem (1998): 158
Authors: Tojyo Y, Tanimura A, Matsumoto Y.
Journal: Biochem Biophys Res Commun (1997): 189
Authors: Garcia-Martin E, Martin-Romero FJ, Gutierrez-Merino C.
Journal: J Neurochem (1995): 2757
Authors: Howarth FC, Singh J, Waring JJ, Hustler BI, Bailey M.
Journal: Magnes Res (1995): 299
Authors: Gunther T, Vormann J, Konstanczak P, Schafer A.
Journal: Biochim Biophys Acta (1994): 281
Application notes
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Evaluation of FLIPR Calcium Assays for Screening GPCR and Calcium Channel Targets
Fluorescent Dye AM Esters
Novel Red Fluorescent Calcium Probes for Functional Analysis of GPCRs and Calcium Channel Targets
FAQ
How do I make an AM ester stock solution?
How do you measure Ca2+ response with calcium indicators like Calbryte 520, potassium salt?
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