Abstract
The water-soluble Cu+chelator bathocuproine disulfonate (BCS) is widely used to quantify Cu+ or detect Cu+ formation in Cu2+-initiated oxidation reactions. The dichlorofluorescin (DCFH) assay is commonly used to monitor free radical reactions, reactive oxygen species (ROS), or reactive nitrogen species (RNS). Upon oxidation the non-fluorescent DCFH is converted into the fluorescent compound dichlorofluorescein (DCF). In the present communication we show that the Cu+ reagent BCS strongly facilitated the oxidation of DCFH in the presence of Cu2+ or Cu+. In contrast, 2,2′-dipyridyl (DP), which is also a Cu+-complexing reagent, but not as well known and therefore not as commonly used as BCS, did not cause any oxidative modification of DCFH in the presence of Cu2+ or Cu+. We therefore recommend that DP should be used instead of BCS to complex Cu+ in reactions which are initiated by Cu2+ and when ROS/RNS are analyzed by the DCFH oxidation assay.
Abbreviations
- BCS:
-
bathocuproine disulfonate
- DCFH:
-
dichlorofluorescin
- DCF:
-
dichlorofluorescein
- DP:
-
dipyridyl
- ROS/RNS:
-
reactive oxygen species/reactive nitrogen species
- DCFDA:
-
2′,7′-dichlorodihydrofluorescin diacetate
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Laggner, H., Hermann, M., Gmeiner, B.M.K. et al. Cu2+ and Cu+ bathocuproine disulfonate complexes promote the oxidation of the ROS-detecting compound dichlorofluorescin (DCFH). Anal Bioanal Chem 385, 959–961 (2006). https://doi.org/10.1007/s00216-006-0498-y
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DOI: https://doi.org/10.1007/s00216-006-0498-y