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

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

Cat. No.: HY-D0940 Purity: 99.81%
Handling Instructions

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

For research use only. We do not sell to patients.

H2DCFDA Chemical Structure

H2DCFDA Chemical Structure

CAS No. : 4091-99-0

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Solution
10 mM * 1 mL in DMSO USD 66 In-stock
Estimated Time of Arrival: December 31
Solid + Solvent
10 mM * 1 mL
ready for reconstitution
USD 66 In-stock
Estimated Time of Arrival: December 31
Solid
50 mg USD 60 In-stock
Estimated Time of Arrival: December 31
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Customer Review

Based on 76 publication(s) in Google Scholar

Top Publications Citing Use of Products

Publications Citing Use of MCE H2DCFDA

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

    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: Curr Biol. 2021 May 18;S0960-9822(21)00607-2.

    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.
    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

    • Customer Review

    Description

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

    In Vitro

    Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
    1. H2DCFDA is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use.
    2. Cells are incubated with 5 µM H2DCFDA 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.
    3. Along with the H2DCFDA probe, if indicated, use ROS-insensitive modification of the fluorescein dye DCFDA as a positive control. 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.

    Molecular Weight

    487.29

    Formula

    C₂₄H₁₆Cl₂O₇

    CAS No.
    Shipping

    Room temperature in continental US; may vary elsewhere.

    Storage

    -20°C, protect from light

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

    Solvent & Solubility
    In Vitro: 

    DMSO : 125 mg/mL (256.52 mM; Need ultrasonic)

    Ethanol : 14.29 mg/mL (29.33 mM; Need ultrasonic)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 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
    10 mM 0.2052 mL 1.0261 mL 2.0522 mL
    *Please refer to the solubility information to select the appropriate solvent.
    In Vivo:
    • 1.

      Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% saline

      Solubility: 2.08 mg/mL (4.27 mM); Suspended solution; Need ultrasonic

    • 2.

      Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in saline)

      Solubility: ≥ 2.08 mg/mL (4.27 mM); Clear solution

    • 3.

      Add each solvent one by one:  10% DMSO    90% corn oil

      Solubility: ≥ 2.08 mg/mL (4.27 mM); Clear solution

    *All of the co-solvents are provided by MCE.
    References
    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.

    References

    Purity: 99.82%

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    Product Name:
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    Cat. No.:
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