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The Journal of Cell Biology, Vol 41, 154-161, Copyright © 1969 by Rockefeller University Press

ARTICLE

ISOLATION AND PROPERTIES OF SECRETORY GRANULES FROM RAT ISLETS OF LANGERHANS

: I. Isolation of a Secretory Granule Fraction



S. L. Howell 1, C. J. Fink 1, and P. E. Lacy 1

1 From the Department of Pathology, Washington University, School of Medicine, St. Louis, Missouri 63110.

Dr. Howell's present address is the School of Biological Sciences, University of Sussex, Brighton, England

A method has been devised for the isolation of a secretory granule fraction from isolated rat islets of Langerhans. The islets were homogenized in buffered sucrose, and the homogenate was separated into nuclear, mitochondrial, secretory granule, and microsomal fractions by differential centrifugation. The secretory granule fraction was purified by differential centrifugation in discontinuous sucrose density gradients. A greater degree of purification could be achieved by the use of two successive gradients of this type, although the final yield was greatly reduced. Biochemical and morphological characterization of the fractions was obtained; the secretory granule fraction contained both insulin and glucagon. The limiting membranes of the granules remained intact and the general appearance of the granules was similar to that seen within the whole islet cells.

Submitted on August 23, 1968
Revised on November 7, 1968


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