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Cal-520N™, potassium salt

Cal-520® provides a robust homogeneous fluorescence-based assay tool for detecting intracellular calcium mobilization. Cal-520® AM is a new fluorogenic calcium-sensitive dye with a significantly improved signal to noise ratio and intracellular retention compared to the existing green calcium indicators (such as Fluo-3 AM and Fluo-4 AM). Cells expressing a GPCR or calcium channel of interest that signals through calcium can be preloaded with Cal-520® AM which can cross cell membrane. Once inside the cell, the lipophilic blocking groups of Cal-520™AM are cleaved by esterases, resulting in a negatively charged fluorescent dye that stays inside cells. Its fluorescence is greatly enhanced upon binding to calcium. When cells stimulated with agonists, the receptor signals the release of intracellular calcium, which significantly increase the fluorescence of Cal-520®. The characteristics of its long wavelength, high sensitivity, and >100 times fluorescence enhancement, make Cal-520® AM an ideal indicator for the measurement of cellular calcium. The high S/N ratio and better intracellular retention make the Cal-520® calcium assay a robust tool for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists. Cal-520® sodium or potassium salt is the hydrolyzed salt of Cal-520® AM in cells. It selectively binds to calcium ion, and has the fluorescence that is strongly dependent on calcium concentration. Compared to other Cal-520® indicators, Cal-520N™ has the lowest affinity to calcium with Kd ~ 90 uM.

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of Cal-520N™, potassium salt to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM99.585 µL497.924 µL995.847 µL4.979 mL9.958 mL
5 mM19.917 µL99.585 µL199.169 µL995.847 µL1.992 mL
10 mM9.958 µL49.792 µL99.585 µL497.924 µL995.847 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum

Citations

View all 93 citations: Citation Explorer
Advances in Two-Photon Scanning and Scanless Microscopy Technologies for Functional Neural Circuit Imaging
Authors: Schultz, Simon R and Copel, undefined and , Caroline S and Foust, Am and a J , undefined and Quicke, Peter and Schuck, Renaud
Journal: Proceedings of the IEEE (2017): 139--157
High-Throughput Phenotyping of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Neurons Using Electric Field Stimulation and High-Speed Fluorescence Imaging
Authors: Daily, Neil J and Du, Zhong-Wei and Wakatsuki, Tetsuro
Journal: ASSAY and Drug Development Technologies (2017)
HTS-Compatible Voltage-and Ca 2+-Sensitive Dye Recordings from hiPSC-Derived Cardiomyocytes Using the Hamamatsu FDSS Systems
Authors: Kettenhofen, Ralf
Journal: Stem Cell-Derived Models in Toxicology (2017): 135--152
Direct measurement of TRPV4 and PIEZO1 activity reveals multiple mechanotransduction pathways in chondrocytes
Authors: Servin-Vences, M Rocio and Moroni, Mirko and Lewin, Gary R and Poole, Kate
Journal: eLife (2017): e21074
Synergistic drug combination effectively blocks Ebola virus infection
Authors: Sun, Wei and He, Shihua and Martínez-Romero, Carles and Kouznetsova, Jennifer and Tawa, Gregory and Xu, Miao and Shinn, Paul and Fisher, Ethan G and Long, Yan and Motabar, Omid and others, undefined
Journal: Antiviral Research (2017): 165--172
Page updated on December 6, 2024

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Catalog Number21147
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Physical properties

Dissociation constant (Kd, nM)90000

Molecular weight

1004.17

Solvent

Water

Spectral properties

Excitation (nm)

492

Emission (nm)

515

Quantum yield

0.751

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200