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RAT ADRENAL DOPAMINE‐β‐HYDROXYLASE PURIFICATION AND IMMUNOLOGIC CHARACTERISTICS
Author(s) -
Grzanna R.,
Coyle J. T.
Publication year - 1976
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1976.tb00313.x
Subject(s) - ouchterlony double immunodiffusion , antiserum , complement fixation test , chemistry , chromatography , immunodiffusion , microbiology and biotechnology , ammonium sulfate precipitation , sephadex , biology , biochemistry , enzyme , antibody , immunology , size exclusion chromatography , serology
— Dopamine‐β‐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.