Left: Western Blot analysis of tyrosine hydroxylase (TH) expression in PC12 cell lysate (Lane 1: 6 ug) and mouse brain (Lane 2: 40 ug). The anti-TH rabbit antibody (1 µg/mL) detects one specific band at ~60 kDa corresponding to the expected molecular weight of TH in both samples. 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-148-50 (1 µg/mL), 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 tuberoinfundibular dopaminergic neurons in rat hypothalamic arcuate nucleus by Immunohistochemistry. Primary antibody: rabbit anti-TH cat# R-148-50 (1:2,000); Secondary antibody: Cy3-conjugated anti-rabbit. Photo courtesy of Dr. Erik Hrabovszky, Hungarian Academy of Sciences, Budapest, Hungary.