Separation of Hormonal Activities in the Anterior Pituitary by Ultracentrifugation

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Abstract
Partial separation of the hormones of the anterior pituitary of beef has been obtained by ultracentrifugation in the Spinco Model L Ultracentrifuge. Fresh and frozen glands were homogenized in 0.25 m sucrose and centrifuged at 500 x g to remove nuclear and cellular debris, at 6600 x g to obtain mitochondria, at 11,150 x g to obtain acidophiles, and at 80,000 x g to obtain basophiles. Each fraction was analyzed for nitrogen, for PNA and for hormonal content. Most of the nitrogen was obtained in the supernatant, most of the PNA in the acidophil granules, and the activity was partially separated. In general the mitochondria contained growth hormones, acidophil granules contained ACTH, supernatant contained FSH, and TSH and LH were found in both acidophil and basophil granules.

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