1. Dye Reagents

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

Cat. No.: HY-D0940 Purity: 99.14%
Handling Instructions

H2DCFDA is a cell-permeable non-fluorescent (but fluorescein-containing) probe.

For research use only. We do not sell to patients.
H2DCFDA Chemical Structure

H2DCFDA Chemical Structure

CAS No. : 4091-99-0

Size Price Stock Quantity
10 mM * 1 mL in DMSO USD 66 In-stock
50 mg USD 60 In-stock
100 mg   Get quote  
200 mg   Get quote  

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Customer Review

  • Biological Activity

  • Protocol

  • Technical Information

  • Purity & Documentation

  • References


H2DCFDA is a cell-permeable non-fluorescent (but fluorescein-containing) probe.

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].

Preparing Stock Solutions
Concentration Volume 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

H2DCFDA is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use[1].
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 fluorescein 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.

Molecular Weight








Powder -20°C 3 years
  4°C 2 years
In solvent -80°C 6 months
  -20°C 1 month

Room temperature in continental US; may vary elsewhere

Solvent & Solubility

DMSO: ≥ 150 mg/mL

* "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

Purity: 99.14%

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