2. Immunology/Inflammation NF-κB Metabolic Enzyme/Protease
  3. Reactive Oxygen Species
  4. H2DCFDA

H2DCFDA  (Synonyms: DCFH-DA; 2',7'-Dichlorodihydrofluorescein diacetate)

目录号: HY-D0940 纯度: 99.81%
COA 产品使用指南

摩尔计算器

H2DCFDA (DCFH-DA) 是一种可渗透细胞的，用于检测细胞内活性氧 (ROS) 的探针 (Ex/Em=488/525 nm)。

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H2DCFDA Chemical Structure

H2DCFDA Chemical Structure

CAS No. : 4091-99-0

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10 mg ￥250
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25 mg ￥370
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50 mg ￥500
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100 mg ￥700
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Customer Review

H2DCFDA 相关抗体

Top Publications Citing Use of Products

MCE 顾客使用本产品发表的 272 篇科研文献

IF

    H2DCFDA purchased from MCE. Usage Cited in: Adv Sci (Weinh). 2022 Mar 10;e2103317.  [Abstract]

    CLSM images of HUVEC cells using DCFH‐DA (0.5 hours) as the fluorescent probe: Control, H2O2, H2O2+10 µg/mL1 PDA NPs, H2O2+40 µg/mL PDA NPs, H2O2+80 µg/mL PDA NPs.

    H2DCFDA purchased from MCE. Usage Cited in: Acta Pharm Sin B. 16 July 2022.

    Treated HL-1 cells are washed three times and incubated with DMEM medium containing H2DCFDA (10 μM; 30 min) at 37 °C. The cells then are washed three times with warm DMEM medium and intracellular ROS production is detected by flow cytometry.

    H2DCFDA purchased from MCE. Usage Cited in: Biomaterials. 2022 Sep 30;290:121842.  [Abstract]

    DCFH-DA (10 μM; 30 min) or C11-BODIPY-labeled cells are substantially increased after treatment with 1 μM FeSO4.

    H2DCFDA purchased from MCE. Usage Cited in: Biomaterials. 2022 May;284:121513.  [Abstract]

    DCFH-DA (10 μM; 30 min). The ROS generation in 4T1 cells and NIH-3T3 cells incubated with various formulations for 4 h observed by CLSM.

    H2DCFDA purchased from MCE. Usage Cited in: Biomaterials. 2022 Jul 21;287:121688.  [Abstract]

    CLSM images of 4T1 tumor cells stained with DCFH-DA (15 min) after different treatments.

    H2DCFDA purchased from MCE. Usage Cited in: J Hazard Mater. 2022 Oct 5;439:129502.  [Abstract]

    Representative DCFH-DA (10 μM; 30 min) staining images (Green) in PBNs exposed PSNPs and/or LPS in vivo and in vitro.

    H2DCFDA purchased from MCE. Usage Cited in: Cell. 2021 Jun 24;184(13):3528-3541.e12.  [Abstract]

    Stained protoplasts with H2DCFDA for the detection of ROS production and PI for PM integrity and tracked the process by live-cell imaging.

    H2DCFDA purchased from MCE. Usage Cited in: ACS Nano. 2021 Oct 26;15(10):16286-16297.  [Abstract]

    CLSM image of 4T1 cells after being treated with MM, MMP, and MMTP; cells are stained with DCFH-DA (20 μM; for 1 h).

    H2DCFDA purchased from MCE. Usage Cited in: Biomaterials. 2021 Oct;277:121103.  [Abstract]

    In vitro experiments are conducted to measure the cellular ROS levels in A549 cells from different treatments via flow cytometry using fluorescent probe H2DCFDA (5 μM; 30 min) to better understand the potential molecular mechanism of SOD@ARA290-HBc on radiation-induced lung injury. H2DCFDA fluorescence intensity in A549 cells significantly increased after irradiation.

    H2DCFDA purchased from MCE. Usage Cited in: Small. 2021 Aug;17(32):e2101368.  [Abstract]

    Laser scanning confocal fluorescence microscopy images of HT-1080 cells treated with different nanodrugs for the detection of intracellular lipid peroxidation and total ROS using C11-BODIPY and DCFH-DA (100 nM; 10 min) as the corresponding fluorescence probes.

    H2DCFDA purchased from MCE. Usage Cited in: Curr Biol. 2021 May 18;S0960-9822(21)00607-2.  [Abstract]

    Co-staining stigmas with H2DCFDA (50 μM; for 1-2 hours) and the cell wall indicator propidium iodide (PI) shows that ROS is located in the cytoplasm of papilla cells, near the periphery of the plasma membrane.

    • 生物活性

    • 实验参考方法

    • 纯度 & 产品资料

    • 参考文献

    生物活性

    H2DCFDA (DCFH-DA) is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS) (Ex/Em=488/525 nm)[1].

    体外研究
    (In Vitro)

    指南 (以下是我们推荐的方案，本方案仅提供指南，应根据您的具体需要进行修改)。
    1. H2DCFDA 溶解在DMSO中得到 10 mM 的储备液，使用前进一步稀释。
    2. 细胞与含5 μM H2DCFDA 的PBS 溶液在37°C 避光孵育30 分钟，然后用0.05% 胰蛋白酶-EDTA 溶液收集细胞，悬浮于新鲜培养基中，立即用流式细胞仪分析。
    3. 如果需要，与 H2DCFDA 探针一起使用荧光素染料 DCFDA 的 ROS 不敏感修饰作为阳性对照。染色程序与 H2DCFDA 相同[1]。

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

    H2DCFDA 相关抗体:
    分子量

    487.29

    Formula

    C24H16Cl2O7
    CAS 号
    性状

    固体

    颜色

    White to off-white

    Emission (Em)

    517
    Excitation (Ex)

    492
    运输条件

    Room temperature in continental US; may vary elsewhere.
    储存方式

    -20°C, protect from light

    *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

    溶解性数据
    In Vitro: 

    DMSO 中的溶解度 : 125 mg/mL (256.52 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)

    Ethanol 中的溶解度 : 20 mg/mL (41.04 mM; 超声助溶)

    配制储备液
    浓度 溶剂体积 质量 1 mg 5 mg 10 mg
    1 mM 2.0522 mL 10.2608 mL 20.5217 mL
    5 mM 0.4104 mL 2.0522 mL 4.1043 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 (protect from 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 (4.27 mM); 悬浊液; 超声助溶

      此方案可获得 2.08 mg/mL的均匀悬浊液，悬浊液可用于口服和腹腔注射。

      以 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% (20% SBE-β-CD in Saline)

      Solubility: ≥ 2.08 mg/mL (4.27 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。

      以 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 20% 的 SBE-β-CD 生理盐水水溶液 中，混合均匀。

      20% SBE-β-CD in Saline 的配制（4°C，储存一周）：2 g SBE-β-CD（磺丁基醚 β-环糊精）粉末定容于 10 mL 的生理盐水中，完全溶解至澄清透明。
    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

    只
    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 (protect from 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: 99.82%

    COA (196 KB) HNMR (228 KB) LCMS (120 KB)

    产品使用指南 (1538 KB)
    染色示例
    参考文献
    Kinase Assay

    ROS Measurements[1]
    For the detection of intracellular ROS level ,ROS-sensitive probe H2DCFDA is used. Adherent cells (ESCs, difESCs, eMSCs, HeLa, U118) are incubated with 5 µM staining solution in PBS in the dark for 30 min at 37°C, then harvested with 0.05% trypsin-EDTA solution, suspended in a fresh medium, and immediately analyzed with flow cytometer. Lymphocytes, both control and PHA-activated, are resuspended in PBS, incubated with 5 µM of H2DCFDA in the dark for 30 min at 37°C, and immediately analyzed. Along with the H2DCFDA probe, if indicated, ROS-insensitive modification of the fluorescent dye DCFDA is used. The staining procedure is the same as for the H2DCFDA[1].

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

    H2DCFDA 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 (protect from 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
    Ethanol / DMSO 1 mM 2.0522 mL 10.2608 mL 20.5217 mL 51.3042 mL
    5 mM 0.4104 mL 2.0522 mL 4.1043 mL 10.2608 mL
    10 mM 0.2052 mL 1.0261 mL 2.0522 mL 5.1304 mL
    15 mM 0.1368 mL 0.6841 mL 1.3681 mL 3.4203 mL
    20 mM 0.1026 mL 0.5130 mL 1.0261 mL 2.5652 mL
    25 mM 0.0821 mL 0.4104 mL 0.8209 mL 2.0522 mL
    30 mM 0.0684 mL 0.3420 mL 0.6841 mL 1.7101 mL
    40 mM 0.0513 mL 0.2565 mL 0.5130 mL 1.2826 mL
    DMSO 50 mM 0.0410 mL 0.2052 mL 0.4104 mL 1.0261 mL
    60 mM 0.0342 mL 0.1710 mL 0.3420 mL 0.8551 mL
    80 mM 0.0257 mL 0.1283 mL 0.2565 mL 0.6413 mL
    100 mM 0.0205 mL 0.1026 mL 0.2052 mL 0.5130 mL
    Help & FAQs
    • Do most proteins show cross-species activity?

      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.

    Keywords:

    H2DCFDA4091-99-0DCFH-DA 2',7'-Dichlorodihydrofluorescein diacetateReactive Oxygen SpeciesInhibitorinhibitorinhibit

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