A rapid procedure for the isolation of bioactive growth hormone.
A two-step isolation method is described for the purification of growth hormone and prolactin from rat pituitaries. Following alkaline extraction, the sample was chromatographed on Sephadex G-75. Growth hormone and prolactin, which eluted in the same gel filtration fraction, were separated on a reversed-phase HPLC column. The isolated proteins migrated to the same position as rat growth hormone and prolactin standards. Further confirmation that the purified proteins were indeed growth hormone and prolactin was provided by immunoblotting using several mammalian antisera and N-terminal amino acid sequencing. The purified rat growth hormone was shown to be biologically active when tested in the hypophysectomized rat body weight gain assay. Although rat prolactin remains to be tested, it seems likely that this hormone would also have biological activity. The present procedure is of general applicability and particularly useful when only small amounts of pituitary glands are available for extraction.
Rand-Weaver
M
M
Laboratory of Molecular Endocrinology, School of Fisheries Sciences, Kitasato University, Iwate, Japan.
Kawauchi
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001835:Body Weight / Q000187:drug effects; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D013006:Growth Hormone / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000737:chemistry / Q000378:metabolism; D011388:Prolactin / Q000302:isolation & purification; D051381:Rats
D011388:Prolactin; D013006:Growth Hormone