Cal Green™ 1, AM [Equivalent to Calcium Green-1, 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) | 190 |
Molecular weight | 1290.96 |
Solvent | DMSO |
Excitation (nm) | 498 |
Emission (nm) | 517 |
Quantum yield | 0.751 |
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 186501-28-0 | Molecular weight 1290.96 | Dissociation constant (Kd, nM) 190 | Excitation (nm) 498 | Emission (nm) 517 | Quantum yield 0.751 |
Platform
Fluorescence microscope
Excitation | FITC |
Emission | FITC |
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 Cal Green™ 1 AM in high-quality, anhydrous DMSO.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Cal Green™ 1 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Cal Green™ 1 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, Cal Green™ 1 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 Cal Green™ 1 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 Cal Green™ 1 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 | 77.462 µL | 387.309 µL | 774.617 µL | 3.873 mL | 7.746 mL |
5 mM | 15.492 µL | 77.462 µL | 154.923 µL | 774.617 µL | 1.549 mL |
10 mM | 7.746 µL | 38.731 µL | 77.462 µL | 387.309 µL | 774.617 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Spectral properties
Excitation (nm) | 498 |
Emission (nm) | 517 |
Quantum yield | 0.751 |
Citations
Authors: Zhang, Nisi and Guo, Yutong and Foiret, Josquin and Tumbale, Spencer K and Paulmurugan, Ramasamy and Ferrara, Katherine W
Journal: STAR protocols (2023): 102723
Authors: Jiang, Xiao-wen and Lu, Hong-yuan and Xu, Zi-Hua and Zhang, Ying-Shi and Zhao, Qing-Chun and others,
Journal: Fitoterapia (2021): 104824
Authors: Sörensen-Zender, Inga and Chen, Rongjun and Song, Rong and David, Sascha and Melk, Anette and Haller, Hermann and Schmitt, Rol and , undefined
Journal: Translational Research (2019)
Authors: Jia, Shang and Ramos-Torres, Karla M and Kolemen, Safacan and Ackerman, Cheri M and Chang, Christopher J
Journal: (2017)
Authors: Jia, Shang and Ramos-Torres, Karla M and Kolemen, Safacan and Ackerman, Cheri M and Chang, Christopher J
Journal: ACS chemical biology (2017): 1844--1852
References
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Journal: Methods Mol Biol (2011): 377
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Journal: Nat Chem Biol (2007): 423
Authors: Lu Y, Paige MF.
Journal: J Fluoresc (2007): 739
Authors: Silei V, Fabrizi C, Venturini G, Tagliavini F, Salmona M, Bugiani O, Lauro GM.
Journal: Brain Res Brain Res Protoc (2000): 132
Authors: Tojyo Y, Tanimura A, Matsumoto Y.
Journal: Biochem Biophys Res Commun (1997): 189
Authors: McPherson DR, McClellan AD, O'Donovan MJ.
Journal: Brain Res Brain Res Protoc (1997): 157
Authors: Askerlund P., undefined
Journal: Plant Physiol (1996): 913
Authors: Spencer CI, Berlin JR.
Journal: Pflugers Arch (1995): 579
Authors: Lloyd QP, Kuhn MA, Gay CV.
Journal: J Biol Chem (1995): 22445
Authors: Rajdev S, Reynolds IJ.
Journal: Neurosci Lett (1993): 149
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
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