CFDA-SE

CFDA-SE（carboxycein diacetate, Succinimidyl Ester）具有细胞膜渗透性和荧光性，在可视化追踪或细胞增殖检测中得到广泛应用。

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10mg

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￥2338.00

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C29H19NO11

货号：ajci12354
CAS：150347-59-4
分子式：C29H19NO11
分子量：557.46
纯度：98%
存储：Store at -20°C
库存：现货

Background:

CFSE is a fluorescent dye which can track the cell division.

CFSE is a fluorescent dye which can track the cell division[1]. To examine the ability of immune cells to migrate into live slices, a single-cell suspension of autologous splenocytes or PBMC is isolated, labeling them with CFSE, and adding them to the top of the slices on day 2 of culture. After 6 additional days of culture, the CFSE-labeling immune cells (green) are found to have migrated throughout the slices. No positive cells are found in the slices without the addition of CFSE-labeling immune cells[2].

参考文献:
[1]. Cinthia C. Stempin, et al. GRAIL and Otubain-1 are Related to T Cell Hyporesponsiveness during Trypanosoma cruzi Infection. PLoS Negl Trop Dis. 2017 Jan; 11(1): e0005307.
[2]. Xiuyun Jiang et al. Long-lived pancreatic ductal adenocarcinoma slice cultures enable precise study of the immune microenvironment. Oncoimmunology. 2017; 6(7): e1333210.

Protocol

Protocol for Cell labeling and counting with CFDA-SE ^[1]:
Cells were counted and stained with CFDA-SE, as follows: 10 mm stock solution of CFDA-SE (Molecular Probes) was prepared in DMSO and aliquots were stored at -20°C in the presence of a desiccant. Harvested cells were washed in HBSS and FBS 1%, resuspended at 20 × 10⁶/mL HBSS with FBS 1%, and stained in a 1:2000 dilution of stock CFDA-SE in HBSS to achieve a final concentration of 2.5 μM (cells were mixed to CFDA-SE 1:1 v/v). The procedure was carried out at room temperature in the dark. At the end of the incubation period (8 min) the reaction was stopped by adding an equal volume of FBS. Cells were gently pelleted at 300 g for 10 min and washed (400 g for 10 min) twice in culture medium Cells were plated and the day after the first flow cytometric acquisition was performed (time point 1). In populations showing a particularly high proliferative rate, two or more generations of cells might be already evident at this point. However, the analysis at the subsequent time points was carried out by comparing the stain distribution with the one described at time point 1. Dead cells were excluded and analysis of acquisition files was performed separately on the two subpopulations. This protocol only provides a guideline, and should be modified according to your specific needs.
References: [1]. Urbani S, Caporale R,et,al. Use of CFDA-SE for evaluating the in vitro proliferation pattern of human mesenchymal stem cells. Cytotherapy. 2006;8(3):243-53. doi: 10.1080/14653240600735834. PMID: 16793733.
Cell experiment^[1]:
Cell lines	Human erythroleukaemic cell line K562, mouse lymphoma cell line YAC-1, human mammary cancer cell line MCF-7 and human melanoma cell line A375
Preparation method	The 5 mM CFDA-SE stock in DMSO was 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. Cells were 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.
Reaction Conditions	1, 1.5, 2, 2.5, 5 and 10 µM CFDA-SE
Applications	CFDA-SE at 2.5 μM stained more than 95% of the cells on all cell lines tested, and dose-dependently increased fluorescence intensity of stained cells. The optimal concentration for K562 and YAC-1 was found to be 2.5 μM, while the optimal concentrations for A375 and MCF-7 were found to be 5 μM and 10 μM, respectively. Within the 6 h experiment period, no cytotoxicity related to CFDA-SE was observed.
Animal experiment [2]:
Animal models	C57BL/6 mice, aged 5 ~ 8 weeks
Preparation method	Injected CFDA-SE into thymic lobe
Dosage form	The concentration of CFDA-SE is 10 μM
Applications	CFDA-SE, at 80 times the concentration used for in vitro labeling, was nontoxic and labeled randomly approximately 15% of thymocytes 24 h after injection. The turnover rate of labeled thymic emigrants in the lymph nodes was in the order of 21 days. Thus, CFDA-SE may serve as a powerful tool in relatively long-term migration studies.
References: [1]: Wang XQ, Duan XM,et,al. Carboxyfluorescein diacetate succinimidyl ester fluorescent dye for cell labeling. Acta Biochim Biophys Sin (Shanghai). 2005 Jun;37(6):379-85. doi: 10.1111/j.1745-7270.2005.00051.x. PMID: 15944752. [2]: Graziano M, St-Pierre Y, et,al. The fate of thymocytes labeled in vivo with CFSE. Exp Cell Res. 1998 Apr 10;240(1):75-85. doi: 10.1006/excr.1997.3900. PMID: 9570923.