Unit

UNIT 3.39 Isolation of Chromaffin Granules

  1. Carl E. Creutz

Published Online: 1 SEP 2010

DOI: 10.1002/0471143030.cb0339s48

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Creutz, C. E. 2010. Isolation of Chromaffin Granules. Current Protocols in Cell Biology. 48:3.39:3.39.1–3.39.10.

Author Information

  1. Department of Pharmacology, University of Virginia, Charlottesville, Virginia

Publication History

  1. Published Online: 1 SEP 2010
  2. Published Print: SEP 2010

Abstract

Adrenal medullary chromaffin granules (dense core secretory vesicles) have been a valuable model system for the study of the proteins and membrane components involved in the process of exocytosis. Because of the abundance of chromaffin granules in a readily available tissue source, bovine adrenal medullae, and their unique sedimentation properties, it is possible to obtain large quantities of highly purified granules and granule membranes in a short period of time. Two protocols are presented here for the isolation of chromaffin granules: a basic protocol based on differential centrifugation in an iso-osmotic medium that yields intact chromaffin granules, and an alternate protocol based on sedimentation through a density step gradient that provides a greater yield of more highly purified chromaffin granules. Since in the latter case the granules cannot be returned to a medium of physiological osmolarity without lysis after purification on the step gradient, the alternate protocol is more useful to obtain the granule membranes or contents for further study. Curr. Protoc. Cell Biol. 48:3.39.1-3.39.10. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • chromaffin granule;
  • chromaffin granule membrane;
  • secretory vesicle;
  • dense core vesicle;
  • adrenal medulla;
  • catecholamine;
  • chromogranin;
  • exocytosis;
  • cytochrome b562;
  • dopamine-β-hydroxylase;
  • bovine