Rabbit polyclonal anti-DMPO antibody was used to detect DMPO-protein-derived nitrone adducts using a standard ELISA as described previously [31 (link)]. In brief, brain homogenates (nigrostriatal region only) were immobilized by incubating reaction mixtures (2.5 mg/well) in 100 mM bicarbonate buffer, pH 9.6, for 90 min at 35°C. The plate was washed once and blocked (4% fish gelatin in PBS) for 90 min at 35°C. Following another wash, rabbit polyclonal anti-DMPO antibody (1:5,000) was added and incubated for 60 min. After incubation, plate wells were washed four times followed by incubation with anti-rabbit, IgG-alkaline phosphatase-conjugated secondary antibody (1:5,000) for 60 min at 35°C. After another four washes, CDP star (25 mM) was added and the chemiluminescent product was detected using a Tecan Spectra Fluor-Plus multifunction plate reader with Xfluor software (Tecan US, Research Triangle Park, NC, USA).