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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


Images

Fluorescent Cell Imaging - Fluo-8 AM, green fluorescent calcium binding dye (AB142773), expandable thumbnail
  • Chemical Structure - Fluo-8 AM, green fluorescent calcium binding dye (AB142773), expandable thumbnail
  • Functional Studies - Fluo-8 AM, green fluorescent calcium binding dye (AB142773), expandable thumbnail
  • Functional Studies - Fluo-8 AM, green fluorescent calcium binding dye (AB142773), expandable thumbnail

Publications

Key facts

CAS number
1345980-40-6
Form
Solid
Molecular weight
1046.94 g/mol
Molecular formula
C50H50N2O23
Nature
Synthetic
Excitation/Emission
Ex: 490nm, Em: 520nm

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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

Key facts

Solubility

Soluble in DMSO.

Biological description

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.

Canonical SMILES
CC(=O)OCOC(=O)CN(CC(=O)OCOC(C)=O)C5CCCCC5OCCOC1CC(CCC1N(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O)C=2C4CCC(CC4OC3=CC(=O)C=CC=23)OCOC(C)=O
InChI
InChI=1S/C50H50N2O23/c1-30(53)65-25-70-37-12-14-39-44(20-37)75-43-19-36(58)11-13-38(43)50(39)35-10-15-41(52(23-48(61)73-28-68-33(4)56)24-49(62)74-29-69-34(5)57)45(18-35)64-17-16-63-42-9-7-6-8-40(42)51(21-46(59)71-26-66-31(2)54)22-47(60)72-27-67-32(3)55/h6-15,18-20H,16-17,21-29H2,1-5H3
InChIKey
BKYHTWFRZPRHNM-UHFFFAOYSA-N
IUPAC name
Bis(acetoxymethyl) 2,2'-((4-(6-(acetoxymethoxy)-3-oxo-3H-xanthen-9-yl)-2-(2-(bis(2-acetoxymethoxy)-2-oxoethyl)amino)phenoxy)ethoxy)phenyl)azanediyl)diacetate
Excitation/Emission
Ex: 490nm, Em: 520nm

Storage

Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
Store under desiccating conditions, The product can be stored for up to 12 months

Notes

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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4 product images

  • Fluorescent Cell Imaging - Fluo-8 AM, green fluorescent calcium binding dye (ab142773), expandable thumbnail

    Fluorescent Cell Imaging - Fluo-8 AM, green fluorescent calcium binding dye (ab142773)

    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.

  • Chemical Structure - Fluo-8 AM, green fluorescent calcium binding dye (ab142773), expandable thumbnail

    Chemical Structure - Fluo-8 AM, green fluorescent calcium binding dye (ab142773)

    2D chemical structure image of ab142773, Fluo-8 AM, green fluorescent calcium binding dye

  • Functional Studies - Fluo-8 AM, green fluorescent calcium binding dye (ab142773), expandable thumbnail
    Cui et al PLoS One. 2013;8(3):e59921. doi: 10.1371/journal.pone.0059921. Epub 2013 Mar 20. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - Fluo-8 AM, green fluorescent calcium binding dye (ab142773)

    FFA treatments increase Ca2+ influx to induce ERS in β-TC3 cells.

    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)

  • Functional Studies - Fluo-8 AM, green fluorescent calcium binding dye (ab142773), expandable thumbnail

    Functional Studies - Fluo-8 AM, green fluorescent calcium binding dye (ab142773)

    Fluo-8 vs Fluo-4 sensitivity to calcium release in HEK-293 cells induced by Carbachol.

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Product protocols

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