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Fluo-3, AM
CAS 121714-22-5
Calcium measurement is critical for numerous biological investigations. Fluorescent probes that show spectral responses upon binding Ca2+ have enabled researchers to investigate changes in intracellular free Ca2+ concentrations by using fluorescence microscopy, flow cytometry, fluorescence spectroscopy and fluorescence microplate readers. Fluo-3 and Rhod-2 are most commonly used among the visible light-excitable calcium indicators. Fluo-3 indicators are widely used in flow cytometry and confocal laser-scanning microscopy. More recently, Fluo-3, AM has been extensively used in cell-based high-throughput screening assays for functional GPCR assays. Fluo-3 is essentially nonfluorescent unless bound to Ca2+ and exhibits a quantum yield at saturating Ca2+ of ~0.14 and a Kd for Ca2+ of 390 nM.
<strong>Effect of increased [Ca<sup>2+</sup>]i on the subcellular localization of CacyBP/SIP in colon cancer SW480 cells.&nbsp;</strong>(A) Effect of different concentrations of ionomycin on the localization of endogenous CacyBP/SIP. Cells were treated with ionomycin for 30 min, followed by immunostaining using anti-CacyBP/SIP, and were imaged with confocal microscopy. CacyBP/SIP was translocated to the perinuclear region in SW480 cells. After stimulation with an increasing amount of ionomycin (0, 1, 2, 5, 10 &mu;mol/L) for 30 min at 37&deg;C, SW480 cells were fixed and immunostained using CacyBP/SIP MAb (panels a, d, g, j, and m), and nuclei were labelled with DAPI (panels b, e, h, k, and n). The merged images are shown in panels c, f, i, l, and o. The scale bar represents 50 &mu;m. (B) The intensity of cytosolic free intracellular Ca<sup>2+</sup> fluorescence in SW480 cells treated with ionomycin (0, 1, 2, 5, 10 &mu;mol/L). The Fluo-3 fluorescence intensity in SW480 cells reached a plateau at 5 &mu;mol/L and 10 &mu;mol/L of ionomycin. SW480 cells were loaded with 20 &mu;mol/L of Fluo-3/AM for 45 min under a confocal microscope (495 nm). The fluorescence was captured every 2 sec and recorded for 3 min. (C) The bar chart shows the intracellular Fluo-3 intensity. Ca<sup>2+</sup> concentration is increased by treatment with 2, 5, and 10 &mu;mol/L of ionomycin (***P&lt;0.001). Source:&nbsp;<strong>The effect of S100A6 on nuclear translocation of CacyBP/SIP in colon cancer cells</strong> by Shanshan Feng et al., <em>PLOS</em>, March 2018.
<strong>Effect of increased [Ca<sup>2+</sup>]i on the subcellular localization of CacyBP/SIP in colon cancer SW480 cells.&nbsp;</strong>(A) Effect of different concentrations of ionomycin on the localization of endogenous CacyBP/SIP. Cells were treated with ionomycin for 30 min, followed by immunostaining using anti-CacyBP/SIP, and were imaged with confocal microscopy. CacyBP/SIP was translocated to the perinuclear region in SW480 cells. After stimulation with an increasing amount of ionomycin (0, 1, 2, 5, 10 &mu;mol/L) for 30 min at 37&deg;C, SW480 cells were fixed and immunostained using CacyBP/SIP MAb (panels a, d, g, j, and m), and nuclei were labelled with DAPI (panels b, e, h, k, and n). The merged images are shown in panels c, f, i, l, and o. The scale bar represents 50 &mu;m. (B) The intensity of cytosolic free intracellular Ca<sup>2+</sup> fluorescence in SW480 cells treated with ionomycin (0, 1, 2, 5, 10 &mu;mol/L). The Fluo-3 fluorescence intensity in SW480 cells reached a plateau at 5 &mu;mol/L and 10 &mu;mol/L of ionomycin. SW480 cells were loaded with 20 &mu;mol/L of Fluo-3/AM for 45 min under a confocal microscope (495 nm). The fluorescence was captured every 2 sec and recorded for 3 min. (C) The bar chart shows the intracellular Fluo-3 intensity. Ca<sup>2+</sup> concentration is increased by treatment with 2, 5, and 10 &mu;mol/L of ionomycin (***P&lt;0.001). Source:&nbsp;<strong>The effect of S100A6 on nuclear translocation of CacyBP/SIP in colon cancer cells</strong> by Shanshan Feng et al., <em>PLOS</em>, March 2018.
protocol
Protocol
sds
SDS
COO
COA
CatalogSize
Price
Quantity
210101 mg
Price
 
Physical properties
Dissociation constant (Kd, nM)390
Molecular weight1129.85
SolventDMSO
Spectral properties
Extinction coefficient (cm -1 M -1)
86,000
1
Excitation (nm)506
Emission (nm)515
Quantum yield
0.15
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
CAS121714-22-5
Instrument settings
Fluorescence microscope
ExcitationFITC
EmissionFITC
Recommended plateBlack wall/clear bottom
Fluorescence microplate reader
Excitation490
Emission525
Cutoff515
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling
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Page updated on October 6, 2025