Cal-630™ AM
Price | |
Catalog Number | |
Availability | In stock |
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) | 792 |
Molecular weight | 1282.89 |
Solvent | DMSO |
Excitation (nm) | 609 |
Emission (nm) | 626 |
Quantum yield | 0.371 |
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 |
Molecular weight 1282.89 | Dissociation constant (Kd, nM) 792 | Excitation (nm) 609 | Emission (nm) 626 | Quantum yield 0.371 |
Platform
Fluorescence microscope
Excitation | Texas Red |
Emission | Texas Red |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 600 |
Emission | 640 |
Cutoff | 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 Cal-630™ AM in anhydrous DMSO.
Note: When reconstituted in DMSO, Cal-630™ AM is a clear, colorless solution.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Cal-630™ AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Cal-630™ 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-630™ 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-630™ 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-630™ 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 2 hours 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 Texas Red filter set or a fluorescence plate reader containing a programmable liquid handling system such as an FDSS, FLIPR, or FlexStation, at Ex/Em = 600/640 nm cutoff 630 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 77.949 µL | 389.745 µL | 779.49 µL | 3.897 mL | 7.795 mL |
5 mM | 15.59 µL | 77.949 µL | 155.898 µL | 779.49 µL | 1.559 mL |
10 mM | 7.795 µL | 38.975 µL | 77.949 µL | 389.745 µL | 779.49 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Spectral properties
Excitation (nm) | 609 |
Emission (nm) | 626 |
Quantum yield | 0.371 |
Product Family
Name | Excitation (nm) | Emission (nm) | Quantum yield |
Cal-590™ AM | 574 | 588 | 0.621 |
Cal-520®, AM | 492 | 515 | 0.751 |
Cal-520FF™, AM | 492 | 515 | 0.751 |
Cal-520N™, AM | 492 | 515 | 0.751 |
Calbryte™ 630 AM | 607 | 624 | - |
Cal-500™ AM | 388 | 482 | 0.481 |
Images
Citations
Authors: Cambra, Hailey M and Tallapragada, Naren P and Mannam, Prabhath and Breault, David T and Klein, Allon M
Journal: Current protocols (2022): e330
Authors: Dickerson, Matthew T and Dadi, Prasanna K and Butterworth, Regan B and Nakhe, Arya Y and Graff, Sarah M and Zaborska, Karolina E and Schaub, Charles M and Jacobson, David A
Journal: The Journal of Physiology (2020): 4887--4905
Authors: Miranda, Caroline
Journal: (2020)
Authors: Liang, Xicai and Yao, Yingjia and Lin, Ying and Kong, Liang and Xiao, Honghe and Shi, Yue and Yang, Jingxian
Journal: Life Sciences (2019)
Authors: Birkner, Antje
Journal: (2019)
Authors: Pioner, Jos{\`e} Manuel and Santini, Lorenzo and Palandri, Chiara and Martella, Daniele and Lupi, Flavia and Langione, Marianna and Querceto, Silvia and Grandinetti, Bruno and Balducci, Valentina and Benzoni, Patrizia and others,
Journal: International journal of molecular sciences (2019): 3799
Authors: Miyazaki, Kenichi and Lisman, John E and Ross, William N
Journal: Frontiers in cellular neuroscience (2018)
Authors: Zhang, Shirley L and Yue, Zhifeng and Arnold, Denice M and Artiushin, Gregory and Sehgal, Amita
Journal: Cell (2018): 130--139
Authors: Zhang, Shirley L and Yue, Zhifeng and Arnold, Denice M and Sehgal, Amita
Journal: bioRxiv (2017): 196956
Authors: Schmunk, Galina
Journal: (2017)
References
Authors: Stamm C, del Nido PJ.
Journal: Thorac Cardiovasc Surg (2004): 127
Authors: Stamm C, Friehs I, Choi YH, Zurakowski D, McGowan FX, del Nido PJ.
Journal: Cardiovasc Res (2003): 695
Authors: Du C, MacGowan GA, Farkas DL, Koretsky AP.
Journal: Biophys J (2001): 549
Authors: Du C, MacGowan GA, Farkas DL, Koretsky AP.
Journal: Cell Calcium (2001): 217
Authors: Lannergren J, Westerblad H, Bruton JD.
Journal: J Muscle Res Cell Motil (2001): 265
Authors: MacGowan GA, Du C, Glonty V, Suhan JP, Koretsky AP, Farkas DL.
Journal: J Biomed Opt (2001): 23
Authors: Muriel MP, Lambeng N, Darios F, Michel PP, Hirsch EC, Agid Y, Ruberg M.
Journal: J Comp Neurol (2000): 297
Authors: Del Nido PJ, Glynn P, Buenaventura P, Salama G, Koretsky AP.
Journal: Am J Physiol (1998): H728
Application notes
Cal-520 ® , Cal-590 ™, and Cal-630™ Calcium Detection Reagents
Calibration Protocol for Fluorescent Calcium Indicators
Novel Red Fluorescent Calcium Probes for Functional Analysis of GPCRs and Calcium Channel Targets
A New Protein Crosslinking Method for Labeling and Modifying Antibodies