Cell permeable analog of FLUO 3. Fluorescent probe for Ca2+. After crossing the membrane, the product is quickly metabolized by cytoplasmic esterases to the membrane impermeant FLUO 3.
Product Details
| Alternative Name: | 1-[Amino-5-(2,7-dichloro-6-acetomethoxy-3-oxo-3H-xanthen-9-yl)phenoxy]-2-(2'-amino-5'-methylphenoxy)ethane-N,N,N',N'-tetraacetic acid, pentaacetoxy-methyl ester |
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| Formula: | C51H50Cl2N2O23 |
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| MW: | 1129.9 |
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| CAS: | 121714-22-5 |
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| Purity: | ≥85% (HPLC) |
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| Identity: | Determined by 1H-NMR and fluorescence. |
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| Appearance: | Red solid. |
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| Solubility: | Soluble in DMSO (0.9mg/ml) and acetonitrile (2mg/ml). |
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| Shipping: | Ambient Temperature |
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| Long Term Storage: | -20°C |
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| Handling: | Protect from light. |
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| Protocol: | General Protocol (for Human T cells)
Reagents:
2mM Fluo 3/AM/DMSO (1mg Fluo 3/AM in 442μl DMSO)
Pluronic F127
Hanks-balanced salt solution (HBSS)
HEPES buffer saline (10mM HEPES, 1mM Na2HPO4, 137mM NaCl, 5mM KCl, 1mM CaCl2, 0.5mM MgCl2, 5mM glucose, 0.1% BSA, pH 7.4)
Protocol:
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Add 16.5mg Pluronic F127 to Fluo 3/AM/DMSO solution. Pluronic F127 prevents aggregation of Fluo 3/AM in HBSS and helps uptake with cells.
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Dilute the Fluo 3/AM solution with HBSS to prepare 4μM Fluo 3/AM working solution.
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Add the Fluo 3/AM working solution to the cells, and incubate at 37°C for 20min.
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Add 5 volumes of HBSS containing 1% fetal calf serum, and continue the incubation for another 40min.
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Wash the cells 3 times with HEPES buffer saline. Then resuspend the cells to prepare 1x105 cells/ml solution using HEPES buffer saline.
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Incubate at 37°C for 10min. Then use the cells for fluorescent calcium ion detection.
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Monitor the fluorescence at 528nm (excitation: 490-500nm)
Cell staining conditions differ by cell types, so it is necessary to optimize the conditions for each experiment. |
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| Regulatory Status: | RUO - Research Use Only |
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Product Literature References
The Us3 Protein of Herpes Simplex Virus 1 Inhibits T Cell Signaling by Confining LAT Activation via TRAF6: Y. Yang, et al.; J. Biol. Chem.
290, 15670 (2015),
Application(s): Cell Culture,
Abstract;
Full Text
Recombinant human tumor necrosis factor alpha induces calcium oscillation and calcium-activated chloride current in human neutrophils. The role of calcium/calmodulin-dependent protein kinase: M.A. Schumann, et al.; J. Biol. Chem.
268, 2134 (1993),
Abstract;
Full Text
Control of mitochondrial matrix calcium: studies using fluo-3 as a fluorescent calcium indicator: A. Saavedra-Molina, et al.; BBRC
167, 148 (1990),
Abstract;
Flow cytometric measurement of cytoplasmic free calcium in human peripheral blood T lymphocytes with fluo-3, a new fluorescent calcium indicator: P.A. Vadenberghe & J.L. Ceuppens; J. Immunol. Meth.
127, 197 (1990),
Abstract;
Glutamate induces calcium waves in cultured astrocytes: long-range glial signaling: A.H. Cornell-Bell, et al.; Science
247, 470 (1990),
Abstract;
Photochemically generated cytosolic calcium pulses and their detection by fluo-3: J.P.Y. Kao, et al.; J. Biol. Chem.
264, 8179 (1989),
Abstract;
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