Fluo-8 AM, a green fluorescent medium affinity calcium-binding dye binds to intracellular calcium (Kd = 390 nM). Fluorescence intensity increases upon Ca2+ binding.
- Cited in over 50 publications
- Cell-permeable
- Soluble in DMSO
- Brighter (1.5x) than Fluo-4 in cellular experiments
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Fluo-8 AM, a green fluorescent medium affinity calcium-binding dye binds to intracellular calcium (Kd = 390 nM). Fluorescence intensity increases upon Ca2+ binding.
- Cited in over 50 publications
- Cell-permeable
- Soluble in DMSO
- Brighter (1.5x) than Fluo-4 in cellular experiments
Soluble in DMSO.
Medium affinity green fluorescent calcium binding dye. Binds to intracellular calcium (Kd = 390 nM). Fluorescence intensity increases upon Ca2+ binding. Cell-permeable.
Fluo-8 (or Fluo-2 Medium Affinity) has been found to be brighter (1.5x) than Fluo-4 in cellular experiments. It offers improved cell loading and Ca2+ 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 loading can be performed at room temperature.
Fluo-8® AM Working Solution: On the day of the experiment, either dissolve Fluo-8® AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature. Prepare a dye working solution of 2 to 20 µM in a buffer of your choice (e.g., Hanks and Hepes buffer) with 0.04% Pluronic® F-127. For most cell lines, Fluo-8® AM at a final concentration of 4-5 μM is recommended. The exact concentration of indicators required for cell loading must be determined empirically.
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U2OS cells were seeded overnight at 40,000 cells per 100 μL per well in a 96-well black wall/clear bottom costar plate. The growth medium was removed, and the cells were incubated with 100 μl of 4 μM Fluo-4 AM or Fluo-8® AM in HHBS at 37 °C, 5% CO2 incubator for 1 hour. The cells were washed twice with 200 μl HHBS, then imaged with a fluorescence microscope using FITC channel.
2D chemical structure image of ab142773, Fluo-8 AM, green fluorescent calcium binding dye
β-TC3 cells were incubated with FFA or BSA for 16 h, and then stimulated with 4 μM thapsigargin for 20 min to activate store-operated Ca2+ entry. Fluorescence densities of Ca2+ change were monitored in Fluo-8/AM-loaded β-TC3 cells after FFA or BSA treatments.
(From Figure 2B of Cui et al)
Fluo-8 vs Fluo-4 sensitivity to calcium release in HEK-293 cells induced by Carbachol.
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