Immunohistochemical Detection of Tyrosine Hydroxylase and Ceruloplasmin

The sections were washed three times by 0.01 M PBS and treated with 0.3% hydrogen peroxide methanol solution for 20 min to suppress endogenous peroxidases activity. After washing thrice in PBS, these sections were treated for 30 min at room temperature with 5% bovine serum albumin for the purpose of reducing nonspecific binding and then incubated overnight with rabbit anti-rat TH (1 : 1,000, GeneTex) [27 (link)] and rabbit anti-rat CP (1 : 1,000, Epitomics) [28 (link)] at 4°C in a humidified chamber. The sections were washed with PBS and incubated with biotinylated goat anti-rabbit IgG (Boster) for 30 min at 37°C, accompanied with enlargement through streptavidin peroxidase for 30 min at the same temperature. Afterwards, sections were rinsed by PBS and coloured with DAB kits for 10 min.