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Cal-520®, sodium 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.

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of Cal-520®, sodium 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 mM118.971 µL594.856 µL1.19 mL5.949 mL11.897 mL
5 mM23.794 µL118.971 µL237.942 µL1.19 mL2.379 mL
10 mM11.897 µL59.486 µL118.971 µL594.856 µL1.19 mL

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

Product family

NameExcitation (nm)Emission (nm)Quantum yield
Cal-520®, potassium salt4925150.751
Cal-590™, sodium salt5745880.621
Cal-630™, sodium salt6096260.371

Citations

View all 94 citations: Citation Explorer
High hydrostatic pressure induces slow contraction in mouse cardiomyocytes
Authors: Yamaguchi, Yohei and Nishiyama, Masayoshi and Kai, Hiroaki and Kaneko, Toshiyuki and Kaihara, Keiko and Iribe, Gentaro and Takai, Akira and Naruse, Keiji and Morimatsu, Masatoshi
Journal: Biophysical Journal (2022)
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
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
Page updated on October 8, 2024

Ordering information

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10x50 ug
1 mg
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Physical properties

Dissociation constant (Kd, nM)320

Molecular weight

840.54

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
ATP-stimulated calcium responses of endogenous P2Y receptor in CHO-K1 cells incubated with Cal-520&trade; AM (red curve), or Fluo-4 AM (blue curve) respectively with (left) or without probenecid (right) under the same conditions. CHO-K1 cells were seeded overnight at 50,000 cells per 100 &micro;L per well in a Costar black wall/clear bottom 96-well plate. 100 &micro;L of 5 &micro;M Fluo-4 AM or Cal 520&trade; AM in HHBS (with or without probenecid) was added into the cells, and the cells were incubated at 37 &deg;C for&nbsp;1 hour. ATP (50 &mu;L/well) was added using FlexSation to achieve the final indicated concentrations.
ATP-stimulated calcium responses of endogenous P2Y receptor in CHO-K1 cells incubated with Cal-520&trade; AM (red curve), or Fluo-4 AM (blue curve) respectively with (left) or without probenecid (right) under the same conditions. CHO-K1 cells were seeded overnight at 50,000 cells per 100 &micro;L per well in a Costar black wall/clear bottom 96-well plate. 100 &micro;L of 5 &micro;M Fluo-4 AM or Cal 520&trade; AM in HHBS (with or without probenecid) was added into the cells, and the cells were incubated at 37 &deg;C for&nbsp;1 hour. ATP (50 &mu;L/well) was added using FlexSation to achieve the final indicated concentrations.
ATP-stimulated calcium responses of endogenous P2Y receptor in CHO-K1 cells incubated with Cal-520&trade; AM (red curve), or Fluo-4 AM (blue curve) respectively with (left) or without probenecid (right) under the same conditions. CHO-K1 cells were seeded overnight at 50,000 cells per 100 &micro;L per well in a Costar black wall/clear bottom 96-well plate. 100 &micro;L of 5 &micro;M Fluo-4 AM or Cal 520&trade; AM in HHBS (with or without probenecid) was added into the cells, and the cells were incubated at 37 &deg;C for&nbsp;1 hour. ATP (50 &mu;L/well) was added using FlexSation to achieve the final indicated concentrations.