Abstract
Publisher Summary The isolation of Golgi apparatus allows a direct approach to the study of the physical, chemical, and enzymatic properties of this cell component provided that the preparations are free of other cell fractions and representative of the state of the Golgi apparatus in situ. The electron microscope has been, and will continue to be, indispensable in the qualitative assay of isolated Golgi apparatus fractions. Quantitative estimates of yield and purity are provided by assays for enzyme activities concentrated in Golgi apparatus and other cell fractions. Centrifugal forces are calculated for the middle of the tube. All solutions are prepared in distilled water. Specific activities of enzymes are given as micromoles of substrate transformed per minute per milligram of protein. The procedures described for isolation of Golgi apparatus from rat liver have been successfully modified for use with rat mammary gland and for mucopolysaccharide-secreting glands of a snail (Helix pomatia). However, no general scheme of fractionation can be given because sedimentation behavior of Golgi apparatus varies from one source to another. The homogenization medium, the method of tissue homogenization, and the sucrose gradient procedure seem to be applicable to most tissues.
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