CFSE

製品コードS8269 バッチS826901

印刷

化学情報

Chemical Structure Synonyms Carboxyfluorescein succinimidyl ester, 5(6)-Carboxyfluorescein diacetate succinimidyl ester, CFDA-SE, 5(6)-CFDA N-succinmidyl ester Storage
(From the date of receipt)		 3 years -20°C(in the dark) powder
化学式

C29H19NO11
分子量 557.46 CAS No. 150347-59-4
Solubility (25°C)* 体外 DMSO 100 mg/mL (179.38 mM)
Water Insoluble
Ethanol Insoluble
体内 （毎回新しく調製した物を用意してください）
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O
2.5mg/ml Taking the 1 mL working solution as an example, add 50 μL of 50 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
Clear solution
5% DMSO 95% Corn oil
5.0mg/ml Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results. 
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

溶剤液（一定の濃度）を調合する

生物活性

製品説明 Carboxyfluorescein succinimidyl ester (CFSE, 5(6)-Carboxyfluorescein diacetate succinimidyl ester, CFDA-SE, 5(6)-CFDA N-succinmidyl ester) is a fluorescent cell staining dye. CFSE is cell permeable and covalently couples, via its succinimidyl group, to intracellular molecules, notably, to intracellular lysine residues and other amine sources.
in vitro

Preparation of CFDA-SE working solution
1.1 Preparation of the stock solution
Dissolve 1 mg of CFDA-SE in 0.1794 mL of DMSO to obtain 10 mM of CFSE.
Note: It is recommended to store the stock solution at -20 ℃ or -80 ℃ away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of CFDA-SE working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 5-10 μM of CFDA-SE working solution.
Note: Please adjust the concentration of CFDA-SE working solution according to the actual situation.
Cell staining
2.1 For suspension cells: Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of CFDA-SE working solution, and then incubate at room temperature for 30 minutes.
2.3 Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.
in vivo

In vivo labeling of cells with CFSE is feasible with virtually no toxic effect on the labeled or adjacent cells. It has advantages in relatively long-term migration studies as demonstrated by the recovery of T-cells in the peripheral lymphoid organs[2].

プロトコル(参考用のみ)

細胞アッセイ 細胞株 human erythroleukaemic cell line K562, mouse lymphoma cell line YAC-1, human mammary cancer cell line MCF-7 and human melanoma cell line A375
濃度 2 µM, 3 µM, 4 µM, 5 µM, 10 µM and 20 µM
反応時間 5, 6, 7, 8, 10 and 15 min
実験の流れ

The 5 mM CFDA-SE stock in DMSO is diluted to different concentrations(2 µM, 3 µM, 4 µM, 5 µM, 10 µM and 20 µM) in PBS with a total volume of 1 ml. After each cell line is harvested and washed three times with PBS, 1×109 cells are added to equal volume of CFDA-SE with different concentrations and incubated at 37°C for 5, 6, 7, 8, 10 and 15 min with agitation. The labeling reaction is stopped for 1 min by adding an equal volume of heat inactivated fetal bovine serum. The CFDA-SE labeled cells are washed twice with PBS and recounted, and the cell concentration is adjusted to 6×104cells/ml in IMDM containing 10% FCS.
動物実験 動物モデル C57Bl/6 mice
投薬量 10 μM
投与方法 injected into thymic lobe

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長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

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