Unit

UNIT 3.35 Isolation of Platelet Granules

  1. Juliane Nießen1,
  2. Gabriele Jedlitschky1,
  3. Andreas Greinacher2,
  4. Heyo K. Kroemer1

Published Online: 1 MAR 2010

DOI: 10.1002/0471143030.cb0335s46

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Nießen, J., Jedlitschky, G., Greinacher, A. and Kroemer, H. K. 2010. Isolation of Platelet Granules. Current Protocols in Cell Biology. 46:3.35:3.35.1–3.35.14.

Author Information

  1. 1

    Department of Pharmacology, Research Center of Pharmacology and Experimental Therapeutics, Ernst-Moritz-Arndt-University, Greifswald, Germany

  2. 2

    Department of Immunology and Transfusion Medicine, Ernst-Moritz-Arndt-University, Greifswald, Germany

Publication History

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

Abstract

Functional analysis of platelet intracellular structures requires isolation and purification of these cellular compartments. With regard to the function of platelets, both, dense (delta) and alpha granules are relevant target structures. However, the availability of sufficient purification protocols for these structures is rather limited. This unit describes two protocols for isolation and purification of platelet granule structures. The Basic Protocol describes a new technique based on immunolabeling with target-specific antibodies followed by magnetic sorting, whereas the Alternate Protocol describes the more traditional procedure based on differential centrifugation and density-based sedimentation. For both methods, the degree of granule purification can be most easily determined by immunoblotting using various antibodies that recognize structure-specific proteins. The immunomagnetic sorting method is especially good for studies requiring highly purified material (e.g., for the identification of specific transporters and receptors). Curr. Protoc. Cell Biol. 46:3.35.1-3.35.14. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • human platelets;
  • subcellular fractionation;
  • magnetic sorting;
  • sucrose density gradient;
  • granule-structures;
  • immunopurification