Synergy httm multi detection microplate reader

ROS production was assessed using the method of Wang & Joseph (1999) (link) with minor modifications. HepG2 cells were plated into black 96-well plates, seeded and treated as previously described. At the end of the treatment, the medium was removed and the cells were washed with PBS and then incubated with 100 mM dichlorofluorescein diacetate (DCF-DA) in PBS for 30 min at 37 °C. The formation of the fluorescent-oxidized derivative of DCF-DA was monitored at emission wavelength of 530 nm and excitation wavelength of 485 nm in a fluorescence multi-detection reader (Synergy HTTM Multi-detection microplate reader; BioTek Instruments Inc., Vermont, USA). ROS production was expressed as unit of fluorescence (UF) produced by DCF-DA/mg of total protein (UF × 10⁴/mg of proteins) (Marabini et al., 2006 (link)).

ROS production was assessed using the method of Wang and Joseph (1999) with minor modifications [19 (link)]. HepG2 cells were plated into black 96-well plates, seeded and treated as previously described. At the end of the treatment, the medium was removed and the cells were washed with PBS and then incubated with 100 mM dichlorofluorescein diacetate (DCF-DA) in PBS for 30 min at 37 °C. The formation of the fluorescent-oxidized derivatives of DCF-DA was monitored at emission wavelength of 530 nm and excitation wavelength of 485 nm in a fluorescence multi-detection reader (Synergy HTTM Multi-detection microplate reader; BioTek Instruments Inc, VT, USA). ROS production was expressed as relative fluorescence unit (RFU) produced by DCF-DA/mg of total protein [20 (link)].