Fura-FF, AM [Fura-2FF, AM] *CAS 348079-12-9*
Price | |
Catalog Number | |
Unit Size | |
Quantity |
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) | 5500 |
Molecular weight | 1023.80 |
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 348079-12-9 | Molecular weight 1023.80 | Dissociation constant (Kd, nM) 5500 | 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 Fura-FF AM in high-quality, anhydrous DMSO.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Fura-FF AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Fura-FF 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, Fura-FF 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-FF 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 Fura-FF 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 | 97.675 µL | 488.377 µL | 976.753 µL | 4.884 mL | 9.768 mL |
5 mM | 19.535 µL | 97.675 µL | 195.351 µL | 976.753 µL | 1.954 mL |
10 mM | 9.768 µL | 48.838 µL | 97.675 µL | 488.377 µL | 976.753 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Product Family
Name | Excitation (nm) | Emission (nm) |
Fura-8FF™, AM | 354 | 524 |
Fura-2, AM *CAS 108964-32-5* | 336 | 505 |
Fura-2, AM *UltraPure Grade* *CAS 108964-32-5* | 336 | 505 |
Fura Red, AM *CAS 149732-62-7* | 435 | 639 |
Fura-8™, AM | 354 | 524 |
Rhod-FF, AM | 553 | 577 |
Fura-10™, AM | 354 | 524 |
Citations
Authors: O’Connor, Thomas N and Kallenbach, Jacob G and Orciuoli, Haley M and Paris, Nicole D and Bachman, John F and Johnston, Carl J and Hernady, Eric and Williams, Jacqueline P and Dirksen, Robert T and Chakkalakal, Joe V
Journal: Skeletal Muscle (2022): 1--15
Authors: Uchimura, Tomoya and Sakurai, Hidetoshi
Journal: Biomedicines (2021): 1589
Authors: Li, Zhi Ying and Jiang, Wen Yi and Cui, Zong Jie
Journal: Photochemical & Photobiological Sciences (2015): 414--428
Authors: Liang, Hui Yuan and Song, Zhi Min and Cui, Zong Jie
Journal: Biochemical and biophysical research communications (2013): 361--367
References
Authors: Shao M, Wang HM, Liu ZH, Shen P, Cai RX.
Journal: Wei Sheng Wu Xue Bao (2005): 805
Authors: McHugh JM, Kenyon JL.
Journal: Am J Physiol Cell Physiol (2004): C342
Authors: Paredes-Gamero EJ, Franca JP, Moraes AA, Aguilar MO, Oshiro ME, Ferreira AT.
Journal: J Fluoresc (2004): 711
Authors: McConnell G, Riis E.
Journal: Phys Med Biol (2004): 4757
Authors: Xu T, Yang W, Huo XL, Song T.
Journal: J Biochem Biophys Methods (2004): 219
Authors: McConnell G, Smith GL, Girkin JM, Gurney AM, Ferguson AI.
Journal: Opt Lett (2003): 1742
Authors: Fischer W, Franke H, Scheibler P, Allgaier C, Illes P.
Journal: Eur J Pharmacol (2002): 53
Authors: Marrelli SP., undefined
Journal: J Neurosci Methods (2000): 145
Authors: Palmer BM, Moore RL.
Journal: Am J Physiol Cell Physiol (2000): C1278
Authors: Berts A, Minneman KP.
Journal: Eur J Pharmacol (2000): 35
Application notes
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
Evaluation of FLIPR Calcium Assays for Screening GPCR and Calcium Channel Targets
Fluorescent Dye AM Esters
FAQ
How do I make an AM ester stock solution?
How do you measure Ca2+ response with calcium indicators like Calbryte 520, potassium salt?
Can I intracellularly measure mitochondria calcium flux and changes in mitochondria membrane potential at the same time?
Do you offer any products for measuring intracellular calcium concentration or movement by flow cytometry?