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

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

Cat. No.: HY-D0940 Purity: 99.82%
Handling Instructions

H2DCFDA is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS).

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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10 mM * 1 mL in DMSO USD 66 In-stock
Estimated Time of Arrival: December 31
50 mg USD 60 In-stock
Estimated Time of Arrival: December 31
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Customer Review

Based on 24 publication(s) in Google Scholar

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H2DCFDA is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS).

In Vitro

Analysis of H2DCFDA probe oxidation by means of flow cytometry technique showed that the basal level of ROS in difESCs is about 6–7 times larger than that in embryonic stem cells (ESCs). Both H2DCFDA and DCFDA probes are non-fluorescent in their initial form but they undergo multistep conversion inside the cell that results in the formation of fluorescent product dichlorofluorescein (DCF). The only difference between these two probes is that the conversion of H2DCFDA involves oxidation. Therefore, fluorescence of the H2DCFDA-treated cells depends on the intracellular ROS level, in contrast to fluorescence associated with the DCFDA probe. Surprisingly, flow cytometry analysis shows that the difference between fluorescence levels of ESCs and difESCs loaded with H2DCFDA is close to the difference between the signals from DCFDA-treated cells. The latter indicates a ROS-independent reason for low fluorescent signal of oxidized H2DCFDA in ESCs[1].

Molecular Weight









Room temperature in continental US; may vary elsewhere.


-20°C, protect from light

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

Solvent & Solubility
In Vitro: 

DMSO : ≥ 150 mg/mL (307.82 mM)

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

H2O : < 0.1 mg/mL (insoluble)

*"≥" means soluble, but saturation unknown.

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.
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.


Purity: 99.82%

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