Cal-520®, 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) | 320 |
Molecular weight | 1102.95 |
Solvent | DMSO |
Excitation (nm) | 492 |
Emission (nm) | 515 |
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 |
Molecular weight 1102.95 | Dissociation constant (Kd, nM) 320 | Excitation (nm) 492 | Emission (nm) 515 | 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-520® AM in high-quality, anhydrous DMSO.
Note: When reconstituted in DMSO, Cal-520® AM is a clear, colorless solution.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Cal-520® AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Cal-520® 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-520® 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-520® 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-520® 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 1 to 2 hours.
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 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 | 90.666 µL | 453.33 µL | 906.659 µL | 4.533 mL | 9.067 mL |
5 mM | 18.133 µL | 90.666 µL | 181.332 µL | 906.659 µL | 1.813 mL |
10 mM | 9.067 µL | 45.333 µL | 90.666 µL | 453.33 µL | 906.659 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Spectrum
Spectral properties
Excitation (nm) | 492 |
Emission (nm) | 515 |
Quantum yield | 0.751 |
Product Family
Name | Excitation (nm) | Emission (nm) | Quantum yield |
Cal-520® maleimide | 492 | 515 | 0.751 |
Cal-520FF™, AM | 492 | 515 | 0.751 |
Cal-520N™, AM | 492 | 515 | 0.751 |
Cal-520® amine | 492 | 515 | 0.751 |
Cal-520® azide | 492 | 515 | 0.751 |
Cal-520® alkyne | 492 | 515 | 0.751 |
Cal-590™ AM | 574 | 588 | 0.621 |
Cal-630™ AM | 609 | 626 | 0.371 |
Calbryte™ 520 AM | 493 | 515 | 0.751 |
Show More (3) |
Images
Citations
Authors: Das, Ritopa and Le, Duong and Kan, Ho-Man and Le, Thinh T and Park, Jinyoung and Nguyen, Thanh D and Lo, Kevin W-H
Journal: Plos one (2024): e0299579
Authors: Song, Jia and Wu, Jiao and Robichaux, Dexter J and Li, Tingting and Wang, Shuyue and Arredondo Sancristobal, Maria J and Dong, Bingning and Dobrev, Dobromir and Karch, Jason and Thomas, Sandhya S and others,
Journal: Cells (2024): 108
Authors: Oropeza-Almaz{\'a}n, Yuriana and Blatter, Lothar A
Journal: Biomolecules (2024): 144
Authors: Yang, Janice and Daily, Neil and Pullinger, Taylor K and Wakatsuki, Tetsuro and Sobie, Eric A
Journal: bioRxiv (2024): 2024--01
Authors: Regeenes, Romario and Rocheleau, Jonathan V
Journal: Lab on a Chip (2024)
Authors: Chen, Jian and Liu, Yanyan and Chen, Feixiang and Guo, Mengnan and Zhou, Jiajia and Fu, Pengfei and Zhang, Xin and Wang, Xueli and Wang, He and Hua, Wei and others,
Journal: Nature Communications (2024): 405
Authors: Li, Pengju and Zhang, Jing and Hayashi, Hidenori and Yue, Jiping and Li, Wen and Yang, Chuanwang and Sun, Changxu and Shi, Jiuyun and Huberman-Shlaes, Judah and Hibino, Narutoshi and others,
Journal: Nature (2024): 1--9
Authors: Wang, Yu-Si and Li, Bai-Yu and Xing, Yin-Fei and Huang, Ji-Cheng and Chen, Zhi-Song and Yue, Liang and Zou, Ying-Gang and Guo, Bin
Journal: Journal of Agricultural and Food Chemistry (2024)
Authors: Chen, Xiaohuan and Bell, Nicole A and Coffman, Breanna L and Rabino, Agustin and Garcia-Mata, Rafael and Yule, David I and Axelrod, Daniel and Smrcka, Alan and Giovannucci, David R and Anantharam, Arun
Journal: bioRxiv (2024): 2024--01
Authors: Zhao, Jin and He, Chengtong and Fan, Xueyu and Wang, Lin and Zhao, Liao and Liu, Hui and Shen, Wujun and Jiang, Sanwei and Pei, Kaixuan and Gao, Jingjing and others,
Journal: Theranostics (2024): 1390
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Application notes
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Cal-520 ® , Cal-590 ™, and Cal-630™ Calcium Detection Reagents