6-Carboxy-H2DCFDA is a chemically reduced, acetylated form of fluorescein used as an indicator for reactive oxygen species (ROS) in living cells. This reagent is not suitable for working with fixed samples.
6-Carboxy-H2DCFDA is a non-fluorescent compound that begins to fluoresce after the cleavage of acetyl groups by cellular esterases and its oxidation by reactive oxygen species inside the cell. The resulting 6-carboxy-2’,7’-dichlorofluorescein has a bright fluorescence in the green channel (absorption maximum at 504 nm, emission maximum at 525 nm), that can be detected using various methods, such as flow cytometry, plate reading, or fluorescent microscopy.
Acetyl groups in 6-carboxy-H2DCFDA increase its lipophilicity and improve the permeability of the indicator through the cell membrane. After deacetylation by cellular esterases, the compound acquires a charge that allows it to be retained inside the cell. This carboxylated H2DCFDA analog has two additional negative charges that impede its leakage out of the cell.
