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Cal-520®, sodium salt

ATP-stimulated calcium responses of endogenous P2Y receptor in CHO-K1 cells incubated with Cal-520™ 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 µL per well in a Costar black wall/clear bottom 96-well plate. 100 µL of 5 µM Fluo-4 AM or Cal 520™ AM in HHBS (with or without probenecid) was added into the cells, and the cells were incubated at 37 °C for 1 hour. ATP (50 μL/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™ 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 µL per well in a Costar black wall/clear bottom 96-well plate. 100 µL of 5 µM Fluo-4 AM or Cal 520™ AM in HHBS (with or without probenecid) was added into the cells, and the cells were incubated at 37 °C for 1 hour. ATP (50 μL/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™ 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 µL per well in a Costar black wall/clear bottom 96-well plate. 100 µL of 5 µM Fluo-4 AM or Cal 520™ AM in HHBS (with or without probenecid) was added into the cells, and the cells were incubated at 37 °C for 1 hour. ATP (50 μL/well) was added using FlexSation to achieve the final indicated concentrations.
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Physical properties
Dissociation constant (Kd, nM)320
Molecular weight840.54
SolventWater
Spectral properties
Excitation (nm)492
Emission (nm)515
Quantum yield0.751
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200
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Show More (77)

OverviewpdfSDSpdfProtocol


Molecular weight
840.54
Dissociation constant (Kd, nM)
320
Excitation (nm)
492
Emission (nm)
515
Quantum yield
0.751
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

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Spectrum


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spectrum

Spectral properties

Excitation (nm)492
Emission (nm)515
Quantum yield0.751

Product Family


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

Images


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 Mart&iacute;nez-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
Properties of synchronous spontaneous Ca2+ transients in the mural cells of rat rectal arterioles
Authors: Mitsui, Retsu and Hashitani, Hikaru
Journal: Pfl&uuml;gers Archiv-European Journal of Physiology (2017): 1--14
A Critical Period for the Rapid Modification of Synaptic Properties at the VPm Relay Synapse
Authors: Pan, Libiao and Yang, Junhua and Yang, Qian and Wang, Xiaomeng and Zhu, Liya and Liu, Yali and Lou, Huifang and Xu, Chou and Shen, Ying and Wang, Hao
Journal: Frontiers in molecular neuroscience (2017)
Retinal origin of direction selectivity in the superior colliculus
Authors: Shi, Xuefeng and Barchini, Jad and Ledesma, Hector Acaron and Koren, David and Jin, Yanjiao and Liu, Xiaorong and Wei, Wei and Cang, Jianhua
Journal: Nature Neuroscience (2017)
Bidirectional communication between sensory neurons and osteoblasts in an in vitro co-culture system
Authors: Kodama, Daisuke and Hirai, Takao and Kondo, Hisataka and Hamamura, Kazunori and Togari, Akifumi
Journal: FEBS Letters (2017)
Calcium transient assays for compound screening with human iPSC-derived cardiomyocytes: Evaluating new tools
Authors: Daily, Neil J and Santos, Radleigh and Vecchi, Joseph and Kemanli, Pinar and Wakatsuki, Tetsuro
Journal: Journal of evolving stem cell research (2017): 1