Oxidative stress is an imbalance between the production of reactive oxygen species (ROS) and cellular antioxidant capacities. As a result, ROS are involved in many diseases.
Measuring ROS levels is therefore a relevant biological target in the search for antioxidant active agents.
The method used to detect intracellular ROS activity is the measurement of 2′,7′-dichlorodihydrofluorescein acetate (H2DCFDA).
Principle of measuring ROS activity levels using the H₂DCFDA probe [Int. J. Mol. Sci. 2020, 21, 1148]
After diffusion into the cell, H2DCFDA is deacetylated by cellular esterases into non-fluorescent H2DCF, which is then oxidized by ROS into fluorescent 2′,7′-dichlorofluorescein (DCF). Thus, the measured fluorescence reflects the level of intracellular ROS activity.
| Format | 96 wells | ||
| Cellular model | Jurkat (ATCC TIB-152) | ||
| Stimulation | TBHP | ||
| Reference molecule | Quercetol | ||
| Incubation | 3H at 37°C under 5% CO2 | ||
| Revelation | H2DCFDA | ||
Notes :
Allow 50 µL at 10 mM or 3 mg of sample to test 5 concentrations (3 points/concentration)
Recommendation :
