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H2DCFDA (DCFH-DA) Catalog No.GC30006

Size Price Stock Qty
10mM*1mLinDMSO
$49.00
In stock
50mg
$45.00
In stock

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Sample solution is provided at 25 µL, 10mM.

Quality Control

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Protocol

Kinase experiment:

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].

References:

[1]. Lyublinskaya OG, et al. Redox environment in stem and differentiated cells: A quantitative approach. Redox Biol. 2017 Aug;12:758-769.

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Chemical Properties

Cas No. 4091-99-0 SDF Download SDF
Synonyms N/A
Chemical Name N/A
Canonical SMILES O=C(O)C1=CC=CC=C1C2C3=C(OC4=C2C=C(Cl)C(OC(C)=O)=C4)C=C(OC(C)=O)C(Cl)=C3
Formula C24H16Cl2O7 M.Wt 487.29
Solubility DMSO : ≥ 150 mg/mL (307.82 mM);Ethanol : 14.29 mg/mL (29.33 mM; Need ultrasonic);H2O : < 0.1 mg/mL (insoluble) Storage Store at -20°C
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request

Background

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

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].

[1]. Lyublinskaya OG, et al. Redox environment in stem and differentiated cells: A quantitative approach. Redox Biol. 2017 Aug;12:758-769.