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Cal-520®, potassium 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 2.5 mm 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 2.5 mm 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.
Ordering information
Price ()
Catalog Number21140
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Physical properties
Dissociation constant (Kd, nM)320
Molecular weight921.08
SolventWater
Spectral properties
Excitation (nm)493
Emission (nm)515
Quantum yield0.751
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


Molecular weight
921.08
Dissociation constant (Kd, nM)
320
Excitation (nm)
493
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®, 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 mM108.568 µL542.841 µL1.086 mL5.428 mL10.857 mL
5 mM21.714 µL108.568 µL217.136 µL1.086 mL2.171 mL
10 mM10.857 µL54.284 µL108.568 µL542.841 µL1.086 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


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)493
Emission (nm)515
Quantum yield0.751

Product family


NameExcitation (nm)Emission (nm)Quantum yield
Mag-520™ potassium salt507531-
Cal-500™, potassium salt3884820.481
Cal-590™, potassium salt5745880.621
Cal-630™, potassium salt6096260.371
Cal-670™, potassium salt667680-
Cal-770™, potassium salt758783-

Citations


View all 99 citations: Citation Explorer
L-type Ca2+ channel recovery from inactivation in rabbit atrial myocytes
Authors: Martinez-Hernandez, Elizabeth and Blatter, Lothar A and Kanaporis, Giedrius
Journal: Physiological Reports (2022): e15222
Dendritic spikes in apical oblique dendrites of cortical layer 5 pyramidal neurons
Authors: Casta{\~n}ares, Michael Lawrence G and Stuart, Greg J and Daria, Vincent R
Journal: bioRxiv (2020)
Multiphoton minimal inertia scanning for fast acquisition of neural activity signals
Authors: Schuck, Renaud and Go, Mary Ann and Garasto, Stefania and Reynolds, Stephanie and Dragotti, Pier Luigi and Schultz, Simon R
Journal: Journal of neural engineering (2018): 025003
Spatio-temporal modulation of light for stimulation and recording of neuronal activity
Authors: Ma, He and Castanares, Michael Lawrence and Daria, Vincent
Journal: (2018): 1072306
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
Interstitial cell modulation of pyeloureteric peristalsis in the mouse renal pelvis examined using FIBSEM tomography and calcium indicators
Authors: Hashitani, Hikaru and Nguyen, Michael J and Noda, Haruka and Mitsui, Retsu and Higashi, Ryuhei and Ohta, Keisuke and Nakamura, Kei-Ichiro and Lang, Richard J
Journal: Pflügers Archiv-European Journal of Physiology (2017): 1--17
Extensive Ca 2+ leak through K4750Q cardiac ryanodine receptors caused by cytosolic and luminal Ca 2+ hypersensitivity
Authors: Uehara, Akira and Murayama, Takashi and Yasukochi, Midori and Fill, Michael and Horie, Minoru and Okamoto, Toru and Matsuura, Yoshiharu and Uehara, Kiyoko and Fujimoto, Takahiro and Sakurai, Takashi and others, undefined
Journal: The Journal of general physiology (2017): 199--218
Synchronicity and Rhythmicity of Purkinje Cell Firing during Generalized Spike-and-Wave Discharges in a Natural Mouse Model of Absence Epilepsy Complex Spike Synchronicity during GSWDs
Authors: Kros, Lieke and Lindeman, S and er , undefined and Eelkman Rooda, Oscar HJ and Murugesan, Pavithra and Bina, Lorenzo and Bosman, Laurens WJ and De Zeeuw, Chris I and Hoebeek, Freek E
Journal: Frontiers in Cellular Neuroscience (2017): 346
Simultaneous Measurement of Neural Activities of Acute Mouse Hippocampal Slices Using Multi-Electrode Array System and Laser Confocal Calcium Imaging
Authors: Hamasaki, Yuuta and Haba, Natsumi and Iwata, Naoki and Uno, Yoshiki and Saito, Minoru
Journal: Journal of Behavioral and Brain Science (2017): 68
Ca 2+ signals initiate at immobile IP 3 receptors adjacent to ER-plasma membrane junctions
Authors: Thillaiappan, Nagendra Babu and Chavda, Alap P and Tovey, Stephen C and Prole, David L and Taylor, Colin W
Journal: Nature Communications (2017): 1505