Fluo-4 AM

Precautions：Fluo-4 AM readily decomposes in water. We recommend aliquoting and sealing this product for the first use, while keeping it dry.The working concentration of the probe, cell volume, incubation temperature, and incubation time need to be optimized in advance for different experiments.This product will solidify and adhere to the interior wall of the vial at low temperatures, such as 4℃. Prior to use, it should be defrosted by incubation in a 20-25℃ water bath. Please store this product away from light to minimize fluorescence quenching.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1.Dilute an appropriate amount of Fluo-4 AM stock solution with PBS to 0.5-5µM working solution.Note: The working solution must be prepared freshly.2.For cultured cells to be tested, remove the culture medium and wash cells three times with PBS or HBSS.Note: The culture medium must be removed thoroughly as the serum in culture medium contains esterases that will cleave Fluo-4 AM into Fluo-4 and phenol red will increase the fluorescence background.3.Add Fluo-4 AM working solution to fully cover cells. 4.Incubate at 20℃-37℃ for 10-60 min, allowing for the loading of Fluo-4 AM into cells.Note: If the optimal incubation conditions has not been determined for a particular experiment, users can start with the incubation at 37℃ for 30 minutes. If the cells die a lot, reduce the incubation time or temperature appropriately. If the fluorescence intensity is too weak, increase the incubation time appropriately.5.Wash cells thrice with PBS or HBSS. After washing, a further incubation of 20-30 min can be performed to ensure sufficient conversion of Fluo-4 AM in cells.6.If required, treat cells with appropriate drugs as desired.7.Examine cells by fluorescence detection instruments such as confocal microscope, multifunctiional plate reader, fluorescence spectrophotometer, or flow cytometry to determine changes in intracellular calcium concentration.Related Products：Cat. No.Product NamePack SizeS1052 Fura-2 AM (Calcium Fluorescent Probe, 2mM)50μlS1056 Fluo-3 AM (Calcium Fluorescent Probe, 5mM)20μlS1060 Fluo-4 AM (Calcium Fluorescent Probe, 2mM)25μlS1672 Ionomycin 50μl ST068 0.5M EGTA pH8.0 20ml

Specification: 2mM