TEFLabs offers the lowest FLUO-8 AM reagent prices available.
TEFLabs will create any packaging size upon request.
- #0801 TEFLabs Fluo-8 (AM) 2 x 50 μg... $15 [2 plates] Add to Cart
- #0803 TEFLabs Fluo-8 (AM) 10 x 50 μg... $65 [10 plates] Add to Cart
- #0805 TEFLabs Fluo-8 (AM) 20 x 50 μg... $130 [20 plates] Add to Cart
- #0811 TEFLabs Fluo-8 (AM) 5 x 1 mg... $495 [100 plates] Add to Cart
Look for our new TEFLabs Assay kits available in late January.
Fluo-8 is the brightest and easiest loading Fluo indicator, and its long wavelength, high sensitivity, affinity, and fluorescence enhancement (>100x) make Fluo-8 an ideal indicator for the measurement of cellular calcium. Fluo-8 is appropriate for use on a wide range of instrumnts including: FLIPR, FDSS, NOVOStar, FlexStation, ViewLux, IN Cell Analyzer, ArrayScan, and others.
Fluo-8 has become the most popular of many names used to market a specific version of the molecule patented and introduced to the world as Fluo-2 in 1988-1989 (US Patent 5049673, 1991). Fluo-8 is Fluo-2 Medium Affinity which replaces the Methyl group of the Fluo-2 High Affinity with a Hydrogen as described below. Since the patent covering the Fluo-8 molecule hs expired, and royalties are no longer due to its inventors, Fluo-8 has increasingly been used under private label as the dye of choice for many of the most popular calcium assay kits. TEFLabs' Fluo-8 offers a brighter, faster, and cheaper direct substitue to Fluo-4 due to its greater fluorescent enhancement, faster and easier room temperature loading, and significantly reduced price point compared to Fluo-4 (as well as other Fluo providers).
The Fluo-8 molecule, is one of the original Fluo series of Ca2+ indicators invented by Drs. Roger Tsien and Akwasi Minta in the 1980s. The Fluo-3 and Fluo-4 molecules have been the most commonly used among the visible light-excitable calcium indicators, due to their early commercialization by Molecular Probes and Invitrogen. However, Fluo-3 AM and Fluo-4 AM are only moderately fluorescent in live cells upon AM hydrolysis, and depending on cell type and application, loading can be difficult to maximize cellular response. Fluo-8 has been found to be much brighter (1.5x) than Fluo-4 in cellular experiments, as shown in Figures 4.11 and 4.12. The increased brightness of Fluo-8 is partly attributable to its superior loading in most cell types.
TEFLabs Fluo-8 (and thus Fluo-2 Medium Affinity) offers improved cell loading and calcium response while maintaining the convenient Fluo-3 and Fluo-4 spectral wavelength of maximum excitation at 490 nm and maximum emission at 520 nm. Fluo-8 loadng can be performed at room temperature while Fluo-3 AM and Fluo-4 AM may require 37 degree C cell loading.
Experimental DATA
Excitation: 488 nm
Emission: 515 nm
kd: 390 nM
Solubility: H20 (salt form) DMSO (AM form)
Molecular Weight: 877 g/mol (salt) 1047 g/mol (AM)
The protocol for using Fluo-8 is the same that is used for Fluo-4.Typical loading protocol for Fluo-8
- Prepare stock solution of 1 – 10 mM Fluo-8 (AM) in DMSO
- Dilute AM stock solution to 1 – 5 μM in aqueous medium and incubate cells for 30 – 60 minutes at room temperature. (If necessary, premix Pluronic F-127 stock solution (15% wt/vol in DMSO) with the desired volume of Fluo-8 (AM) stock solution before dispersal into aqueous medium.)
- Wash/transfer cells into fresh medium for experimentation (for “wash-free” assays, washing may be omitted).
Table 4.6 TEFLabs Fluo-8(Fluo-2 MedAff) versus Fluo-4 comparison
Table 4.6 shows a comparison of Fluo-8 (TEFlabs Fluo-2 MedAff) and Fluo-4 properties. the pKa of Fluo-8is slightly higher than Fluo-4. Although a lower pKa was preferred when the Fluo dyes were invented, experimental data has established that Fluo-8’s pKa is not a limitation.
Figure. 4.11 FLUO-2,3, 4,8 brightness comparison
Figure. 4.12 FLUO-2, 3, 4, 8 cell comparison
Table 4.7 Fluo-2 MedAff/Fluo-8 properties
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