UNIT 8.1B Purification of Organelles from Mammalian Cells
Published Online: 1 NOV 2003
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Castle, J. D. 2003. Purification of Organelles from Mammalian Cells. Current Protocols in Immunology. 56:I:8.1B:8.1B.1–8.1B.57.
- Published Online: 1 NOV 2003
- Published Print: AUG 2003
Organelle purification procedures capitalize on the differences in size, density, and (occasionally) surface charge density of individual types of organelles. Most fractionation procedures that are based on centrifugation involve some combination of procedures that distinguish both size and density. Initially, a homogenate is prepared in isoosmotic (or slightly hyperosmotic) sucrose or some other predominantly nonelectrolyte medium. A wide range of procedures have been used to fractionate tissue homogenates. The protocols in this unit emphasize different fractionation techniques that have been used for rat liver, an abundant tissue that has been a favorite of many investigators and has served as the source of many organelle preparations of excellent purity. For selected procedures, examples have been given using other tissue sources (e.g., glandular tissues that maintain protein storage granules for regulated secretion) or, where particularly favorable, cultured cells.