Unit

UNIT 8.1B Purification of Organelles from Mammalian Cells

  1. J. David Castle

Published Online: 1 NOV 2003

DOI: 10.1002/0471142735.im0801bs56

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Castle, J. D. 2003. Purification of Organelles from Mammalian Cells. Current Protocols in Immunology. 56:8.1B.1–8.1B.57.

Author Information

  1. University of Virginia, Charlottesville, Virginia

Publication History

  1. Published Online: 1 NOV 2003
  2. Published Print: AUG 2003

Abstract

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.