Simple and accurate assay for tyrosine hydroxylase from bovine retina by high-performance liquid chromatography with fluorescence detection.
We present a simple and sensitive assay for tyrosine hydroxylase (TH) using high-performance liquid chromatography (HPLC) with a fluorescence detector which enabled us to study bovine retinal TH of a small quantity in crude retinal samples. The detection limit with this method was 1 pmol of dihydroxyphenylalanine (DOPA) enzymatically produced. Retinal TH exhibited the maximum activity at pH 6 to 6.3, and almost none at physiological pH of above 7, a tendency similar to that reported for striatal TH.
Mandai
M
M
Department of Ophthalmology, Faculty of Medicine, Kyoto University, Japan.
Iwaki
M
M
Honda
Y
Y
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004295:Dihydroxyphenylalanine / Q000032:analysis; D004795:Enzyme Stability; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D015203:Reproducibility of Results; D012160:Retina / Q000201:enzymology*; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis*
D004295:Dihydroxyphenylalanine; D014446:Tyrosine 3-Monooxygenase