Howell S L, Fink C J, Lacy P E
J Cell Biol. 1969 Apr;41(1):154-61. doi: 10.1083/jcb.41.1.154.
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.
已设计出一种从分离的大鼠胰岛中分离分泌颗粒组分的方法。将胰岛在缓冲蔗糖中匀浆，然后通过差速离心将匀浆液分离成核、线粒体、分泌颗粒和微粒体组分。通过在不连续蔗糖密度梯度中进行差速离心来纯化分泌颗粒组分。使用这种类型的两个连续梯度可实现更高程度的纯化，尽管最终产量会大大降低。获得了各组分的生化和形态学特征；分泌颗粒组分同时含有胰岛素和胰高血糖素。颗粒的界膜保持完整，颗粒的总体外观与整个胰岛细胞内所见相似。
