title: Rat adrenal dopamine-beta-hydroxylase: purification and immunologic characteristics.
abstract: Dopamine-beta-hydroxylase (DBH) was purified from rat adrenal medulla by a series of steps including sedimentation of membranes, extraction with n-butanol, ammonium sulfate fractionation, gel chromatography and ion-exchange chromatography. Disk gel electrophoresis revealed two protein bands, both of which were active. Antiserum was prepared against homogeneously purified bovine adrenal and rat adrenal DBH; Ouchterlony immunodiffusion, enzyme neutralization and complement fixation tests demonstrated that the respective homologous antisera were monospecific and of high titer. Antiserum to bovine DBH was only 2- to 3-fold more potent than pre-immune serum in inhibition of rat DBH activity. Complement fixation tests demonstrate that antiserum to bovine DBH has a 25,000-fold lower immunoreactivity with rat DBH than with bovine DBH.
authors.affiliation: Departments of Pharmacology and Experimental Therapeutics and Psychiatry and the Behavioral Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: R
authors.initials: R
authors.lastname: Grzanna
authors.forename: J T
authors.initials: JT
authors.lastname: Coyle
mesh_terms: D000311:Adrenal Glands / Q000737:chemistry* / Q000201:enzymology*; D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D002845:Chromatography; D003168:Complement Fixation Tests; D004299:Dopamine beta-Hydroxylase / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005779:Immunodiffusion; D009928:Organ Specificity; D051381:Rats; D013552:Swine
chemical_list: D004299:Dopamine beta-Hydroxylase