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  3. CFDA-SE

CFDA-SE  (Synonyms: 5(6)-Carboxyfluorescein diacetate succinimidyl ester; 5(6)-CFDA N-succinmidyl ester)

目录号: HY-D0938 纯度: 99.01%
COA 产品使用指南

CFDA-SE 是一种可穿透细胞膜的荧光染料,其能与胞内细胞骨架蛋白中的游离胺基反应,最终形成具有荧光的蛋白复合物。该染料进入细胞后主要定位于细胞膜、细胞质和细胞核,在细胞核的染色效果最强。CFDA-SE 染料可用于检测细胞增殖,CFDA-SE 可随着细胞的分裂增殖而被子代细胞均匀继承,其含量的衰减与细胞分裂次数成正比,该荧光信号可在 488 nm 的激发光下进行检测,并可通过流式细胞仪进行分析。

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CFDA-SE Chemical Structure

CFDA-SE Chemical Structure

CAS No. : 150347-59-4

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Top Publications Citing Use of Products

MCE 顾客使用本产品发表的 52 篇科研文献

IF
Proliferation Assay

    CFDA-SE purchased from MCE. Usage Cited in: Proc Natl Acad Sci U S A. 2022 Aug 9;119(32):e2201899119.  [Abstract]

    Flow cytometry analysis of CD4+ T cell number (B), proliferation rate (C), and representative images of CFSE-based proliferation assay (D) of CD4+ T cells cocultured with AT2 cells from healthy, LLC-bearing GPX3fl/fl, or GPX3cKO mice respectively.

    CFDA-SE purchased from MCE. Usage Cited in: Cancer Lett. 2022 May 17;541:215746.  [Abstract]

    PBMCs are labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) (2.5 μM). The proliferation of T cells is assessed by CFSE intensity using flow cytometry.

    CFDA-SE purchased from MCE. Usage Cited in: OncoImmunology. 2022 Feb 1;11(1):2032918.  [Abstract]

    Bone marrow cells are incubated with 5 μΜ CFSE (5, 6- carboxyfluorescein diacetate) for 10 min at 37°C.

    CFDA-SE purchased from MCE. Usage Cited in: Theranostics. 2021 Feb 20;11(9):4251-4261.  [Abstract]

    Mouse MC38 cells are washed with PBS and labeled with 1 μM CFDA-SE in PBS for 30 minutes at room temperature.

    CFDA-SE purchased from MCE. Usage Cited in: J Cell Mol Med. 2021 Jul;25(14):7052-7065.  [Abstract]

    THP-1 cells are stained with 1 μM carbox fluorescenceindiacetate succinimidyl ester (CFSE) in 0.1% BSA for 10 minutes at 37°C and ended with PBS for 10 minutes.

    CFDA-SE purchased from MCE. Usage Cited in: Mol Nutr Food Res. 2022 Jan;66(2):e2100619.  [Abstract]

    CFDA-SE (10 μM) is added to bacterial suspension and then incubated at 37°C in the dark for 20 min. The four strains are labeled with fluorescent probe CFDA-SE and were gavaged to baby rats to evaluate their colonization ability. Compared to the control group, the CFDA-SE fluorescence is present in rats gavaged with the four strains.
    • 生物活性

    • 实验参考方法

    • 纯度 & 产品资料

    • 参考文献

    生物活性

    CFDA-SE is a fluorescent dye that can penetrate the cell membrane. It can react with the free amine group in the cytoskeleton protein inside the cell, and finally form a protein complex with fluorescence. After entering the cell, CFDA-SE locates in the cell membrane, cytoplasm and nucleus, and the fluorescence staining is strongest in the nucleus[1]. CFDA-SE dye can be uniformly inherited by the cells with cell division and proliferation, and its attenuation is proportional to the number of cell divisions. This phenomenon can be detected and analyzed by flow cytometry under the excitation light of 488 nm, and can be used to detect the proliferation of cells[1].

    体外研究
    (In Vitro)

    CFDA-SE 工作液的配制
    1.1 储备液的配制
    取 1 mg CFDA-SE 溶于 0.1794 mL DMSO 中,得到 10 mM CFDA-SE。
    注:原液建议 -20℃ 或 -80℃ 避光保存,避免反复冻融。
    1.2 CFDA-SE 工作液的配制
    在无血清细胞培养中稀释原液
    注意:请根据实际情况调整 CFDA-SE 工作液的浓度。
    细胞染色
    2.1 悬浮细胞:4℃,1000 g 离心 3-5 分钟,弃上清。PBS 洗涤 2 次,每次 5 分钟。
    贴壁细胞:弃去细胞培养基,加入胰蛋白酶解离细胞,制成单细胞悬液。4℃ 1000 g 离心 3-5 分钟,弃上清。PBS 洗涤 2 次,每次 5 分钟。
    2.2 加入 1 mL CFDA-SE 工作液,室温孵育 30 分钟。
    2.3 4℃,400 g 离心 3-4 分钟
    2.4 PBS 洗涤 2 次,每次 5 分钟。
    2.5 用无血清细胞培养基或 PBS 重悬细胞,荧光显微镜或流式细胞仪检测。

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    分子量

    557.46

    Formula

    C58H38N2O22

    CAS 号
    性状

    固体

    颜色

    Off-white to yellow

    Emission (Em)

    515

    Excitation (Ex)

    485

    中文名称

    5(6)-羧基二乙酸荧光素琥珀酰亚胺酯

    运输条件

    Room temperature in continental US; may vary elsewhere.

    储存方式

    -20°C, sealed storage, away from moisture and light

    *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

    溶解性数据
    In Vitro: 

    DMSO 中的溶解度 : 50 mg/mL (89.69 mM; 超声助溶; 吸湿的 DMSO 对产品的溶解度有显著影响,请使用新开封的 DMSO)

    配制储备液
    浓度 溶剂体积 质量 1 mg 5 mg 10 mg
    1 mM 1.7939 mL 8.9693 mL 17.9385 mL
    5 mM 0.3588 mL 1.7939 mL 3.5877 mL
    查看完整储备液配制表

    * 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效
    储备液的保存方式和期限:-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C储存时,请在6个月内使用,-20°C储存时,请在1个月内使用。

    • 摩尔计算器

    • 稀释计算器

    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    质量
    =
    浓度
    ×
    体积
    ×
    分子量 *

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    浓度 (start)

    C1

    ×
    体积 (start)

    V1

    =
    浓度 (final)

    C2

    ×
    体积 (final)

    V2

    In Vivo:

    请根据您的 实验动物和给药方式 选择适当的溶解方案。

    以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液,再依次添加助溶剂:
    ——为保证实验结果的可靠性,澄清的储备液可以根据储存条件,适当保存;体内实验的工作液,建议您现用现配,当天使用
    以下溶剂前显示的百分比是指该溶剂在您配制终溶液中的体积占比;如在配制过程中出现沉淀、析出现象,可以通过加热和/或超声的方式助溶

    • 方案 一

      请依序添加每种溶剂: 10% DMSO    40% PEG300    5% Tween-80    45% Saline

      Solubility: ≥ 2.08 mg/mL (3.73 mM); 澄清溶液

      此方案可获得 ≥ 2.08 mg/mL(饱和度未知)的澄清溶液。

      1 mL 工作液为例,取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中,混合均匀;再向上述体系中加入 50 μL Tween-80,混合均匀;然后再继续加入 450 μL 生理盐水 定容至 1 mL

      生理盐水的配制:将 0.9 g 氯化钠,溶解于 ddH₂O 并定容至 100 mL,可以得到澄清透明的生理盐水溶液。
    • 方案 二

      请依序添加每种溶剂: 10% DMSO    90% Corn Oil

      Solubility: ≥ 2.08 mg/mL (3.73 mM); 澄清溶液

      此方案可获得 ≥ 2.08 mg/mL(饱和度未知)的澄清溶液,此方案实验周期在半个月以上的动物实验酌情使用。

      1 mL 工作液为例,取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中,混合均匀。

    动物溶解方案计算器
    请输入动物实验的基本信息:

    给药剂量

    mg/kg

    动物的平均体重

    g

    每只动物的给药体积

    μL

    动物数量

    由于实验过程有损耗,建议您多配一只动物的量
    请输入您的动物体内配方组成:
    %
    DMSO +
    +
    %
    Tween-80 +
    %
    Saline
    如果您的动物是免疫缺陷鼠或者体弱鼠,建议 DMSO 中的在最后工作液体系中的占比尽量不超过 2%。
    方案所需 助溶剂 包括:DMSO ,均可在 MCE 网站选购。 Tween 80,均可在 MCE 网站选购。
    计算结果
    工作液所需浓度 : mg/mL
    储备液配制方法 : mg 药物溶于 μL  DMSO(母液浓度为 mg/mL)。

    *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

    您所需的储备液浓度超过该产品的实测溶解度,以下方案仅供参考,如有需要,请与 MCE 中国技术支持联系。
    动物实验体内工作液的配制方法 : 取 μL DMSO 储备液,加入 μL  μL ,混合均匀至澄清,再加 μL Tween 80,混合均匀至澄清,再加 μL 生理盐水
    连续给药周期超过半月以上,请谨慎选择该方案。
    请确保第一步储备液溶解至澄清状态,从左到右依次添加助溶剂。您可采用超声加热 (超声清洗仪,建议频次 20-40 kHz),涡旋吹打等方式辅助溶解。
    纯度 & 产品资料

    纯度: 99.01%

    染色示例
    参考文献
    Cell Assay
    [2]

    Single-cell suspensions of splenocytes (or PBMC) are stained with 1 µM CFSE in PBS for 9 min at 37°C, combined with 20 mL of 10% FBS RPMI-1640 medium at RT for 2 min, centrifuged, washed, and counted. 2×106 CFSE-labeling splenocytes are added to the top of the slices in 24-well membrane culture insert[2].

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    参考文献

    CFDA-SE 相关分类

    完整储备液配制表

    * 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效
    储备液的保存方式和期限:-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C储存时,请在6个月内使用,-20°C储存时,请在1个月内使用。

    可选溶剂 浓度 溶剂体积 质量 1 mg 5 mg 10 mg 25 mg
    DMSO 1 mM 1.7939 mL 8.9693 mL 17.9385 mL 44.8463 mL
    5 mM 0.3588 mL 1.7939 mL 3.5877 mL 8.9693 mL
    10 mM 0.1794 mL 0.8969 mL 1.7939 mL 4.4846 mL
    15 mM 0.1196 mL 0.5980 mL 1.1959 mL 2.9898 mL
    20 mM 0.0897 mL 0.4485 mL 0.8969 mL 2.2423 mL
    25 mM 0.0718 mL 0.3588 mL 0.7175 mL 1.7939 mL
    30 mM 0.0598 mL 0.2990 mL 0.5980 mL 1.4949 mL
    40 mM 0.0448 mL 0.2242 mL 0.4485 mL 1.1212 mL
    50 mM 0.0359 mL 0.1794 mL 0.3588 mL 0.8969 mL
    60 mM 0.0299 mL 0.1495 mL 0.2990 mL 0.7474 mL
    80 mM 0.0224 mL 0.1121 mL 0.2242 mL 0.5606 mL
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      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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