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Cal-590™ AM
Calcium measurement is critical for numerous biological investigations. Fluorescent probes that show spectral responses upon binding calcium have enabled researchers to investigate changes in intracellular free calcium concentrations by using fluorescence microscopy, flow cytometry, fluorescence spectroscopy and fluorescence microplate readers. Rhod-2 is most commonly used among the red fluorescent calcium indicators. However, Rhod-2 AM is only moderately fluorescent in live cells upon esterase hydrolysis, and has very small cellular calcium responses. Cal-590™ has been developed to improve Rhod-2 cell loading and calcium response while maintaining the spectral wavelength of Rhod-2, making it compatible with TRITC/Cy3® filter set. In CHO and HEK cells Cal-590™ AM has cellular calcium response that is much more sensitive than Rhod-2 AM. The spectra of Cal-590 is well separated from those of FITC, Alexa Fluor® 488 and GFP, making it an ideal calcium probe for multiplexing intracellular assays with GFP cell lines or FITC/Alexa Fluor® 488 labeled antibodies.
ATP-stimulated calcium response of endogenous P2Y receptor in CHO-K1 cells incubated with Cal-590® AM or Rhod-2 AM under the same conditions. CHO-K1 cells were seeded overnight at 50,000 cells per 100 µL per well in a 96-well black wall/clear bottom Costar plate. 100 µL of 5 µg/mL Cal-590® AM or Rhod-2 AM with 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 by FlexStation (Molecular Devices) to achieve the final indicated concentrations.
ATP-stimulated calcium response of endogenous P2Y receptor in CHO-K1 cells incubated with Cal-590® AM or Rhod-2 AM under the same conditions. CHO-K1 cells were seeded overnight at 50,000 cells per 100 µL per well in a 96-well black wall/clear bottom Costar plate. 100 µL of 5 µg/mL Cal-590® AM or Rhod-2 AM with 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 by FlexStation (Molecular Devices) to achieve the final indicated concentrations.
protocol
Protocol
COO
COA
CatalogSize
Price
Quantity
205105x50 ug
Price
2051110x50 ug
Price
205121 mg
Price
 
Physical properties
Dissociation constant (Kd, nM)561
Molecular weight1266.81
SolventDMSO
Spectral properties
Extinction coefficient (cm -1 M -1)
78000
Excitation (nm)574
Emission (nm)588
Quantum yield
0.62
1
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
Instrument settings
Fluorescence microscope
ExcitationTRITC/Cy3
EmissionTRITC/Cy3
Recommended plateBlack wall/clear bottom
Fluorescence microplate reader
Excitation540
Emission590
Cutoff570
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling
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Page updated on October 6, 2025