Multiskan fc microtiter plate reader

The NADH-ubiquinone oxidoreductase activity assay was performed on cell membranes isolated from bovine heart tissue. For this purpose, membrane homogenates (0.1 mg/mL) were incubated in the presence of 0.35 mM NADH and 0.5 mg/mL NBT in phosphate buffer (5 mM; pH 7.4) for 3 h at 25 °C with increasing concentrations (from 0.1 nM to 1 mM) of drugs and compounds with antipsychotic, anticholinergic, narcotic, and analgesic properties (olanzapine, clozapine, desclozapine, fluphenazine, pentazocine, PB 28, DXT, DMT, donepezil, BD 1047, PPCC, and NE 100) in the presence and absence of 50 µM dUQ. The reaction started by the addition of membrane homogenates, and NBT oxidation was measured every 5 min spectrophotometrically at 595 nM in a Multiskan FC microtiter plate reader (Thermo Scientific^®, Waltham, MA, USA). Biochemical data on NADH oxidoreductase-evoked superoxide production were presented as a percentage of basal activity in the absence of the drug under study, with or without decylubiquinone. Every CMMA had three replicates of each tissue, and all the experiments were performed in duplicate.