FLUOFORTE^(R) Calcium assay kit  
FluoForte (商標名)

FLUOFORTE® Calcium assay kit image％％FLUOFORTE^(R) dye increases in fluorescent intensity when bound to calcium.

FLUOFORTE® Calcium assay kit Excitation and emission％％Excitation (solid line) and emission (dotted line) spectra of FLUOFORTE^(R) dye.

FLUOFORTE® Calcium assay kit 17_Fig1_webimage_01％％FLUOFORTE^(R) exhibits significantly brighter fluorescence intensity than Fluo-4 AM and Fluo-3 AM. Evaluation of relative dye fluorescence. U2OS cells were seeded overnight, growth medium was removed, and cells were incubated with 100 µl of FLUOFORTE^(R) AM, Fluo-3 AM, or Fluo-4 AM in HHBS at 37 °C, 5% CO2 incubator for 1 hour. The cells were washed twice with 200 µl HHBS, and ATP (20 µL/well) was added to achieve concentrations of 200 nM with dye efflux inhibitor. Cells were then immediately imaged with a fluorescence microscope (Olympus IX71) using FITC channel.

FLUOFORTE® Calcium assay kit 17_Fig2_webimage％％Obtain robust signal intensity. ATP dose response curves in CHO-M1 cells, expressing P2Y endogenous receptors. CHO cells were seeded overnight at 40,000 cells per 100 µl per well in a 96-well black wall/clear bottom microplate. The cells were incubated with 100 µl of Life Technologies’ Fluo-4 Direct™ kit, Molecular Devices’ Calcium 4 kit (both based upon manufacturer’s protocol) or FLUOFORTE^(R) dye. ATP (20µl/well) was added by FlexStation to achieve the final indicated concentrations. Comparable ATP EC50 values were obtained using all three dyes, while FLUOFORTE^(R) generated the highest intensity signal.