Left: Western Blot analysis of tyrosine hydroxylase (TH) expression in PC12 cell lysate (6 ug protein/lane). The anti-TH rabbit antibody (1:300) detects one specific band at ~60 kDa corresponding to the expected molecular weight of TH. Method: SDS Page: denaturing and reducing, 4-12% Bis-Tris gel; Transfer: Tris-Glycine (Towbin's buffer) with 20% methanol; Membrane: PVDF (0.45 um); Blocking: 5% skim milk in TBST, 1 hr at RT; Primary antibody: R-118-100 (1:300), overnight at 2-8°C; Secondary antibody: donkey anti-rabbit (1:25,000), 1 hr at RT; Detection: Chemiluminescence. Right: Detection of TH-immunoreactivity in dopaminergic neurons in the rat zona incerta in formalin-fixed floated cryostat section by Immunohistochemistry. TH was visualized with the rabbit polyclonal antiserum (R-118-100; 1:100,000) using the biotinylated secondary antibody-ABC method and nickel-diaminobenzidine chromogen. Photo courtesy of Dr. Erik Hrabovszky, Hungarian Academy of Sciences, Budapest, Hungary.