English
中文
CAS:121714-22-5
Appearance:Orange solid
Storage:Powder : -20℃, 2 years;In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:HPLC≥93%
Manual Download
Fluo-3, AM is one of the most commonly used fluorescent probes to detect intracellular calcium ion concentration. After it penetrates the cell membrane and enters the cell, it is shear by the intracellular esterase to form Fluo-3, which is retained in the cell. Fluo-3 is almost non-fluorescent when it exists in the form of free ligand, but when it combines with intracellular calcium ions, it can produce strong fluorescence, and the maximum excitation wavelength is 506nm. The maximum emission wavelength is 526nm. In practice, the recommended excitation wavelength is about 488nm and the emission wavelength is 525-530nm. Changes in intracellular calcium concentration can be detected using laser confocal microscopy or flow cytometry.
Usage (for reference only) :
Fluo-3 AM solution was diluted with HBSS to prepare 4-5μM of Fluo-3 AM working solution.
Fluo-3, AM working solution was added to cells and cultured at 37℃ for 20 minutes.
Five times the volume of HBSS containing 1% fetal bovine serum was added, and the culture was continued for 40 minutes.
The cells were washed with HEPES buffer saline for 3 times, and then the cells were re-suspended with HEPES buffer saline to produce 1×105 cells/mL solution.
The cells were cultured at 37℃ for 10 minutes, and then fluorescent calcium ion detection was performed. The excitation wavelength is 506nm and the emission wavelength is 526nm.
* The conditions for labeling vary depending on the cell type, so please determine the best conditions before each experiment.
Note:
1. If the medium containing serum is used, the lipase in the serum will decompose the AM body, thereby reducing the effect of Fluo-3 and AM entering the cell.
2, the medium containing phenol red will make the background value slightly higher, so before adding the working liquid, the medium residue should be removed as far as possible.
3. Pluronic F127 can prevent Fluo-3, and AM can polymerize in HBSS and help FluO-3 enter cells. Therefore, an appropriate amount of Pluronic F127 solution can be added to Fluo-3,AM solution, and it is not recommended to store Fluo-3,AM solution in Pluronic F127 for a long time.
4. Fluorescent dyes have extraction problems, please pay attention to avoid light to slow down fluorescence extraction.
Examples of using this product（for reference only）
frozen section（5 μM Fluo-3 AM ，30 min at 37 ℃ ）:
Fresh brain specimens were cut into 5 μm sections using a frozen tissue slicer. Specimens were incubated in Hank's balanced salt solution containing 5 μM Fluo-3 AM (Solarbio, China) for approximately 30 min at 37 ℃ and then photographed using a fluorescence microscope. The average fluorescence intensity of the PFC was calculated by Imagepro-Plus software
Reference：
Chen C, Chen W, Nong Z, Nie Y, Chen X, Pan X, Guo Y, Yao M, Deng W. Hyperbaric oxygen alleviated cognitive impairments in mice induced by repeated cerebral ischemia-reperfusion injury via inhibition of autophagy. Life Sci. 2020 Jan 15;241:117170. doi: 10.1016/j.lfs.2019.117170. Epub 2019 Dec 12. PMID: 31838137.
