Preparations enriched in apparently intact secretory vesicles were isolated from homogenates of lactating rat and bovine mammary tissue by differential and density gradient centrifugation in isoosmotic media. Morphologically, these preparations consisted nearly entirely of vesicles of varying sizes, at least some of which containedcasein micelles. Endoplasmic reticulum vesicles, Golgi apparatus cisterna and dictyosomes, mitochondria, peroxisomes, lysosomes, and nuclei were not observed in secretory vesicle-rich fractions. Vesicle preparations were enriched in lactose relative to total membrane fractions from mammary gland. (11, 12). The pellets that collected in centrifuge tubes during secretory vesicle isolation on Ficoll gradients were collected and analyzed for lactose content and will be referred to as the "vesicle-depleted particulate fraction." Total particulate fractions were obtained by centrifugation of clarified homogenates at 120,000 X g (maximum) for 60 min at 2'. Analytical Methods. Protein was determined with the Folin phenol reagent with bovine serum albumin as standard (27).
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