Fluo-3 AM Documentation

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Fluo-3 AM Product Specifications

Fluo-3 AM is a cell-permeable ester commonly used in cell-based experiments to detect changes in intracellular calcium levels.

Molecular Weight

1129 g/mol

Kd

390 nM

Odor

None

Fire and Explosion Hazards

None

Toxicology Data

Not known

Handling and Storage

Store at -20°C. Protect from light and moisture

Shelf Life

Valid for six months after delivery, if stored properly

TLC

Solvent

Ethyl Acetate / Hexane 2:1

Rf

0.4

HPLC

Column

C18

Detector Settings

254 nm, 371 nm

Purity

> 95%

Absorbance Spectrum

Solvent

Ethyl Acetate

Absorbance max

460 nm

Fluorescence Spectrum

Fluoresces after hydrolyzed by esterases in the cell.

Excitation max

506 nm

Emission max

525 nm

1H NMR

All relevant peaks present

Solvent

Deuterated chloroform

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Fluo-3 AM Background

Fluo-4_AM_Theoretical_Background

Most useful and inexpensive lasers operate in the green part of the visible spectrum and that has made green-emitting fluorescent calcium indicators very useful for confocal microscopy, flow cytometry, and screening. The popular green-emitting fluorescent calcium indicators for screening are Fluo-2, Fluo-3, Fluo-4, and Fluo-8. They are all derived from the original Fluo compound invented by Minta and Tsien (1989).

Fluo-3 has been used to image the spatial dynamics of Ca2+ signaling, in flow cytometry experiments involving photoactivation of caged chelators, second messengers, and neurotransmitters, and for cell-based pharmacological screening. Fluo-4 is an analog of Fluo-3 with the two chlorine substituents replaced by fluorines, which results in increased fluorescence excitation at 488 nm and consequently higher fluorescence signal levels. Cells may be loaded with the AM ester forms of these calcium indicators by adding the dissolved indicator directly to dishes containing cultured cells.

The patent for Fluo-2, Fluo-3, and Fluo-4 have expired so Ion Indicators offers these at reasonable prices.

Dye Characteristic Comparison

Reagents for viability and Toxicity Assays

MW (g/mol)

Excitation (nm)

Emission (nm)

Kd

Extinction

Solubilty

Fluo-2 AM

1061

N/A

N/A

N/A

26,000

DMSO

Fluo-2 K+ Salt

891

490

494

230

80,000

H2O

Fluo-3 AM

1130

N/A

N/A

N/A

26,000

DMSO

Fluo-3 K+ Salt

960

506

515

390

80,000

H2O

Fluo-4 AM

1097

N/A

N/A

N/A

26,000

DMSO

Fluo-4 K+ Salt

927

490

526

345

80,000

H2O

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