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Calbryte™ 630 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. x-Rhod-1 is commonly used as a red fluorescent calcium indicator. However, x-Rhod-1 is only moderately fluorescent in live cells upon esterase hydrolysis, and has very small cellular calcium responses. Calbryte™ 630 has been developed to improve x-Rhod-1 cell loading and calcium response while maintaining the spectral wavelength of x-Rhod-1, making it compatible with Texas Red® filter set. In CHO and HEK cells Cal-630™ AM has cellular calcium response that is much more sensitive than x-Rhod-1. The spectra of Calbryte™ 630 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. Calbryte™ 630 is a new generation of red fluorescent indicators for the measurement of intracellular calcium. Its greatly improved signal/background ratio and intracellular retention properties make Calbryte™ 630 AM the most robust deep red fluorescent indicator for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists in live cells. Like other dye AM cell loading, Calbryte™ 630 AM ester is non-fluorescent and once gets inside the cell, it is hydrolyzed by intracellular esterase and gets activated. The activated indicator is a polar molecule that is no longer capable of freely diffusing through cell membrane, essentially trapped inside cells.
Graph illustrates signal-to-noise (SNR) x 100%. ATP dose response was measured in CHO-K1 cells with Calbryte™ 630 AM. CHO-K1 cells were seeded overnight at 50,000 cells/100 µL/well in a 96-well black wall/clear bottom costar plate. 100 µL of 10 µg/ml Calbryte™ 630 AM in HH Buffer with probenecid was added and incubated for 60 min at 37°C. Dye loading solution was then removed and replaced with 200 µL HH Buffer/well. ATP (50 µL/well) was added by FlexStation 3 to achieve the final indicated concentrations.
Graph illustrates signal-to-noise (SNR) x 100%. ATP dose response was measured in CHO-K1 cells with Calbryte™ 630 AM. CHO-K1 cells were seeded overnight at 50,000 cells/100 µL/well in a 96-well black wall/clear bottom costar plate. 100 µL of 10 µg/ml Calbryte™ 630 AM in HH Buffer with probenecid was added and incubated for 60 min at 37°C. Dye loading solution was then removed and replaced with 200 µL HH Buffer/well. ATP (50 µL/well) was added by FlexStation 3 to achieve the final indicated concentrations.
protocol
Protocol
COO
COA
CatalogSize
Price
Quantity
207202x50 ug
Price
2072110x50 ug
Price
207221 mg
Price
 
Physical properties
Dissociation constant (Kd, nM)1200
Molecular weight1234.84
SolventDMSO
Spectral properties
Excitation (nm)607
Emission (nm)624
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
Flow cytometer
Excitation640 nm laser
Emission660/20 nm filter
Instrument specification(s)APC channel
Fluorescence microscope
ExcitationTexas Red
EmissionTexas Red
Recommended plateBlack wall/clear bottom
Fluorescence microplate reader
Excitation600
Emission640
Cutoff630
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling
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Page updated on October 13, 2025