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FLUO 3/AM

Fluorescent probe for Ca2+. Cell permeable.
 
ALX-620-003-M001 1 mg 274.00 USD
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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 Specification

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
 
Formula:C51H50Cl2N2O23
 
MW:1129.9
 
CAS:121714-22-5
 
Purity:≥85% (HPLC)
 
Identity:Determined by 1H-NMR and fluorescence.
 
Appearance:Red solid.
 
Solubility:Soluble in DMSO (0.9mg/ml) and acetonitrile (2mg/ml).
 
Shipping:Ambient
 
Long Term Storage:-20°C
 
Handling:Protect from light.
 
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:

  1. 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.
  2. Dilute the Fluo 3/AM solution with HBSS to prepare 4μM Fluo 3/AM working solution.
  3. Add the Fluo 3/AM working solution to the cells, and incubate at 37°C for 20min.
  4. Add 5 volumes of HBSS containing 1% fetal calf serum, and continue the incubation for another 40min.
  5. Wash the cells 3 times with HEPES buffer saline. Then resuspend the cells to prepare 1x105 cells/ml solution using HEPES buffer saline.
  6. Incubate at 37°C for 10min. Then use the cells for fluorescent calcium ion detection.
  7. 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.

 
ALX-620-003
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ALX-620-003

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; Full Text

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