H2DCFDA is a cell-permeable fluorogenic probe that is useful for the detection of reactive oxygen species (ROS) and the nitric oxide (NO) in neutrophils and macrophages and for the determination of the overall oxidative stress in toxicological phenomena.

Store at -20°C in dark.

Protocol for ROS measurements H2DCFDA is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use. Flow cytometry analysis Adherent cells are incubated with 5 µM H2DCFDA 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 (ex: 485/20, em: 530/30). Confocal microscopy analysis Coverslips with cells are placed in the 5 μM H2DCFDA staining solution in PBS for 60 min at 37 °C in the dark, then washed and imaged with a confocal laser-scanning microscope. {{Lyublinskaya OG, Ivanova JS, Pugovkina NA, Kozhukharova IV, Kovaleva ZV, Shatrova AN, Aksenov ND, Zenin VV, Kaulin YA, Gamaley IA, Nikolsky NN. Redox environment in stem and differentiated cells: A quantitative approach. Redox Biol. 2017 Aug;12:758-769. doi: 10.1016/j.redox.2017.04.016. Epub 2017 Apr 11.|https://pubmed.ncbi.nlm.nih.gov/28426982/}}

H2DCFDA is a cell-permeable fluorogenic probe that is useful for the detection of reactive oxygen species (ROS) and the nitric oxide (NO) in neutrophils and macrophages and for the determination of the overall oxidative stress in toxicological phenomena. Excitation wavelength ~490 nm, emission ~520 nm.