Calbryte™ 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) | 1200 |
Molecular weight | 1090.90 |
Solvent | DMSO |
Excitation (nm) | 493 |
Emission (nm) | 515 |
Quantum yield | 0.751 |
Certificate of Origin | Download PDF |
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 1090.90 | Dissociation constant (Kd, nM) 1200 | Excitation (nm) 493 | Emission (nm) 515 | Quantum yield 0.751 |
Platform
Flow cytometer
Excitation | 488 nm laser |
Emission | 530/30 nm filter |
Instrument specification(s) | FITC channel |
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 Calbryte™ 520 AM in anhydrous DMSO.
Note: When reconstituted in DMSO, Calbryte™ 520 AM is a clear, colorless solution.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Calbryte™ 520 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Calbryte™ 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, Calbryte™ 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 Calbryte™ 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 Calbryte™ 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 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 | 91.667 µL | 458.337 µL | 916.674 µL | 4.583 mL | 9.167 mL |
5 mM | 18.333 µL | 91.667 µL | 183.335 µL | 916.674 µL | 1.833 mL |
10 mM | 9.167 µL | 45.834 µL | 91.667 µL | 458.337 µL | 916.674 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Spectral properties
Excitation (nm) | 493 |
Emission (nm) | 515 |
Quantum yield | 0.751 |
Product Family
Name | Excitation (nm) | Emission (nm) |
Calbryte™ 590 AM | 581 | 593 |
Calbryte™ 630 AM | 607 | 624 |
Calbryte™-520L AM | 493 | 515 |
Calbryte™-520XL AM | 493 | 515 |
Cal-520®, AM | 492 | 515 |
Mag-520™ AM | 506 | 525 |
SoNa™ 520 AM | 491 | 511 |
Images
Citations
Authors: Dehio, Philippe and Michard, C{\'e}line and Yam-Puc, Juan Carlos and Mart{\'\i} i L{\'\i}ndez, Adri{\`a}-Arnau and Fabre, Lucien and Schaefer, Thorsten and Wymann, Matthias P and Okkenhaug, Klaus and Soldati, Thierry and Mehling, Matthias and others,
Journal: bioRxiv (2024): 2024--01
Authors: Sharma, Sadhana and Wahl, Devin and Risen, Sydney and Gilberto, Vincenzo S and Chatterjee, Anushree and Moreno, Julie and LaRocca, Thomas and Nagpal, Prashant
Journal: bioRxiv (2024): 2024--03
Authors: Maltsev, Anna V and Barlas, Yasir and Hazan, Adina T and Zhang, Rui and Goldhaber, Joshua I
Journal: Biophysical Journal (2024): 457a
Authors: Panzer, Julia K and Garcia, Pablo A and Pugliese, Alberto
Journal: JoVE (Journal of Visualized Experiments) (2024): e66468
Authors: Hoven, Darius and Inaoka, Misaki and McCoy, Reece and Withers, Aimee and Owens, R{\'o}is{\'\i}n M and Malliaras, George G
Journal: MRS Communications (2024): 1--6
Authors: Acreman, Samuel and Ma, Jinfang and Denwood, Geoffrey and Gao, Rui and Tarasov, Andrei and Rorsman, Patrik and Zhang, Quan
Journal: iScience (2024)
Authors: Atif, Abdul Raouf and Aramesh, Morteza and Carter, Sarah-Sophia and Tenje, Maria and Mestres, Gemma
Journal: Journal of Materials Science: Materials in Medicine (2024): 2
Authors: Li, Jiaojiao and Zhao, Qin and Wang, Can and Fu, Liangliang and Zhao, Zifan and Tang, Ziqiao and Yin, Chenghu and Wang, Min and Xia, Haibin and others,
Journal: Acta Biomaterialia (2024)
Authors: Huang, Wen and Yang, Shuo and Deng, Minying and Luo, Rongkui and Liang, Huaiyu and Shen, Yanyan and Yang, Biyu and Xu, Chen and Hou, Yingyong
Journal: Functional \& Integrative Genomics (2024): 1--14
References
Authors: Wu, Yanjiao and Xu, Xiaoli and Ma, Lunkun and Yi, Qian and Sun, Weichao and Tang, Liling
Journal: The International Journal of Biochemistry & Cell Biology (2017)
Authors: Lu, Jiang and Yao, Xue-qin and Luo, Xin and Wang, Yu and Chung, Sookja Kim and Tang, He-xin and Cheung, Chi Wai and Wang, Xian-yu and Meng, Chen and Li, Qing and others, undefined
Journal: Neural Regeneration Research (2017): 945
Authors: Yang, Gang and Xiao, Zhenghua and Ren, Xiaomei and Long, Haiyan and Ma, Kunlong and Qian, Hong and Guo, Yingqiang
Journal: Scientific Reports (2017): 41781
Authors: Liu, Jia and Du, Qing and Zhu, He and Li, Yu and Liu, Maodong and Yu, Shoushui and Wang, Shilei
Journal: Int J Clin Exp Med (2017): 6861--6868
Authors: Sun, Xia and Lin, Yi and Huang, Qiansheng and Shi, Junpeng and Qiu, Ling and Kang, Mei and Chen, Yajie and Fang, Chao and Ye, Ting and Dong, Sijun
Journal: Journal of cellular and molecular medicine (2015): 581--594
Authors: Zhao, Lantao and Li, Shuhong and Wang, Shilei and Yu, Ning and Liu, Jia
Journal: Biochemical and biophysical research communications (2015): 537--542
Authors: Liang, Nan and Wang, Peng and Wang, Shilei and Li, Shuhong and Li, Yu and Wang, Jinying and Wang, Min
Journal: Journal of Neural Transmission (2014): 593--600
Authors: Li, Qing and Lu, Jiang and Wang, Xianyu and others, undefined
Journal: Neural regeneration research (2014): 2002
Authors: Peng, Xu-Dong and Zhao, Gui-Qiu and Lin, Jing and Jiang, Nan and Xu, Qiang and Zhu, Cheng-Cheng and Qu, Jain-Qiu and Cong, Lin and Li, Hui
Journal: International journal of ophthalmology (2014): 441--447
Authors: Hagiyama, M and Inoue, T and Furuno, T and Iino, T and Itami, S and Nakanishi, M and Asada, H and Hosokawa, Y and Ito, A
Journal: British Journal of Dermatology (2013): 771--778
Application notes
FAQ
Do you offer any products for measuring intracellular calcium concentration or movement by flow cytometry?
What is fluorescence quantum yield?
Why is my Fluo-4 signal not at a useful magnitude?
Are there any calcium indicators that don't require probenecid (PBC)?