Fura Red, AM *CAS 149732-62-7*
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Dissociation constant (Kd, nM) | 400 |
Molecular weight | 1088.99 |
Solvent | DMSO |
Excitation (nm) | 435 |
Emission (nm) | 639 |
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 | ![]() ![]() |
CAS 149732-62-7 | Molecular weight 1088.99 | Dissociation constant (Kd, nM) 400 | Excitation (nm) 435 | Emission (nm) 639 |
Platform
Fluorescence microplate reader
Excitation | 435, 470 |
Emission | 630, 650 |
Cutoff | Ex/Em = 435/630, cutoff 610. Ex/Em = 470/650, cut off 630 |
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 Red AM in high-quality, anhydrous DMSO.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Fura Red AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Fura Red 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 Red 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 Red 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 Red 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 monitor fluorescence intensity using a fluorescence plate reader, which contains a programmable liquid handling system such as a FlexStation, at Ex/Em1 = 435/630 nm cutoff 610 nm and Ex/Em2 = 470/650 nm cutoff 630 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 91.828 µL | 459.141 µL | 918.282 µL | 4.591 mL | 9.183 mL |
5 mM | 18.366 µL | 91.828 µL | 183.656 µL | 918.282 µL | 1.837 mL |
10 mM | 9.183 µL | 45.914 µL | 91.828 µL | 459.141 µL | 918.282 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-FF, AM [Fura-2FF, AM] *CAS 348079-12-9* | 336 | 505 |
Fura-8™, AM | 354 | 524 |
Calcein Red™ AM | 562 | 576 |
Fura-10™, AM | 354 | 524 |
Citations
Authors: Golosova, Daria and Levchenko, Vladislav and Kravtsova, Olha and Palygin, Oleg and Staruschenko, Alexander
Journal: Scientific reports (2022): 1--12
Authors: Ye, Zhongde and Shen, Yi and Jin, Ke and Qiu, Jingtao and Hu, Bin and Jadhav, Rohit R and Sheth, Khushboo and Weyand, Cornelia M and Goronzy, J{\"o}rg J
Journal: Nature communications (2021): 1--17
Authors: Cameron, Breanne Ashleigh
Journal: (2021)
Authors: Golosova, Daria and Palygin, Oleg and Bohovyk, Ruslan and Klemens, Christine A and Levchenko, Vladislav and Spires, Denisha R and Isaeva, Elena and El-Meanawy, Ashraf and Staruschenko, Alexander
Journal: Life science alliance (2020)
Authors: Cameron, Breanne A and Stoyek, Matthew R and Bak, Jessi J and Quinn, T Alexander
Journal: bioRxiv (2020)
Authors: Lee, Christine SM and Huguenin, Yoann and Pillois, Xavier and Moulieras, Mikeldi and Marcy, Ella and Whittaker, Shane and Chen, Vivien MY and Fiore, Mathieu
Journal: Research and Practice in Thrombosis and Haemostasis
References
Authors: Wendt ER, Ferry H, Greaves DR, Keshav S.
Journal: PLoS One (2015): e0119532
Authors: Bailey S, Macardle PJ.
Journal: J Immunol Methods (2006): 220
Authors: Walczysko P, Wagner E, Albrechtova JT.
Journal: Cell Calcium (2000): 23
Authors: Su ZL, Li N, Sun YR, Yang J, Wang IM, Jiang SC.
Journal: Shi Yan Sheng Wu Xue Bao (1998): 323
Authors: Blackwood AM, Sagnella GA, Mark and u ND, MacGregor GA.
Journal: J Hum Hypertens (1997): 601
Authors: Wu Y, Clusin WT.
Journal: Am J Physiol (1997): H2161
Authors: Floto RA, Mahaut-Smith MP, Somasundaram B, Allen JM.
Journal: Cell Calcium (1995): 377
Authors: Schild D, Jung A, Schultens HA.
Journal: Cell Calcium (1994): 341
Authors: Novak EJ, Rabinovitch PS.
Journal: Cytometry (1994): 135
Authors: Lew VL, Etzion Z, Bookchin RM, daCosta R, Vaananen H, Sassaroli M, Eisinger J.
Journal: Biochim Biophys Acta (1993): 152
Application notes
A Comparison of Fluorescent Red Calcium Indicators for Detecting Intracellular Calcium Mobilization in CHO Cells
A Meta-Analysis of Common Calcium Indicators
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A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
FAQ
Are there any calcium indicators that don't require probenecid (PBC)?
Are there upgraded trypan blue derivatives for cell viability testing?
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?