Primary anti th antibody mouse monoclonal

Briefly, to block endogenous peroxidase activity, brain sections were treated with 0.6% H₂O₂ in Tris-buffered saline (TBS; pH 7.5), blocked in TBS-TS for 30 min, and incubated with primary anti-TH antibody (mouse monoclonal; Chemicon, Temecula, CA, USA) in TBS-TS at 4 °C. Sections were further processed using appropriate biotinylated secondary goat anti-mouse IgG antibodies (Vector Laboratories; Burlingame, CA, USA) at room temperature for 3 h, incubated in ABC solution (ABC reagent Elite Kit, Vector Laboratories; Burlingame, CA, USA) at room temperature for 1 h, and developed with diaminobenzidine (DAB) solution. Images were obtained using a Nikon ECLIPSE TE 2000-U microscope (Nikon, Tokyo, Japan). Densitometric analysis in STR was performed using FluorChem SP software (Alpha Innotech, San Leandro, CA, USA) and TH+ neurons were counted in entire extents of the SN by the same blinded investigator.

Briefly, to block endogenous peroxidase activity, brain sections were treated with 0.6% H₂O₂ in Tris-buffered saline (TBS; pH 7.5), blocked in TBS-TS for 30 min, and incubated with primary anti-TH antibody (mouse monoclonal; Chemicon, Rolling Meadows, IL, USA) in TBS-TS at 4 °C. The sections were then exposed to appropriate biotinylated secondary goat anti-mouse IgG antibodies (Vector Laboratories, San Francisco, CA, USA) at room temperature for 3 h, incubated in ABC solution (ABC reagent Elite Kit, Vector Laboratories) at room temperature for 1 h, and developed using diaminobenzidine (DAB) solution. Images were obtained using a Nikon ECLIPSE TE 2000-U microscope (Nikon, Tokyo, Japan). Densitometric analysis of TH expression intensity in STR (five to six sections per mouse) were performed using a FluorChem SP (Alpha Innotech, San Leandro, CA, USA), and the numbers of TH positive dopaminergic neurons in SN were counted.