Unit

UNIT 9.27 Whole Blood Processing for Measurement of Signaling Proteins by Flow Cytometry

  1. Sue Chow1,
  2. David Hedley1,
  3. T. Vincent Shankey2

Published Online: 1 OCT 2008

DOI: 10.1002/0471142956.cy0927s46

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Chow, S., Hedley, D. and Shankey, T. V. 2008. Whole Blood Processing for Measurement of Signaling Proteins by Flow Cytometry. Current Protocols in Cytometry. 46:9.27:9.27.1–9.27.19.

Author Information

  1. 1

    Division of Applied Molecular Oncology, Ontario Cancer Institute, Princess Margaret Hospital, Toronto, Ontario, Canada

  2. 2

    Advanced Technology Center, Beckman Coulter, Inc., Miami, Florida

Publication History

  1. Published Online: 1 OCT 2008
  2. Published Print: OCT 2008

Abstract

Signal transduction pathways link external stimuli with cellular responses, which normally regulate cell proliferation, death, and differentiation. The study of signal transduction was revolutionized through the development of phospho-specific antibodies that recognize proteins only when they are phosphorylated at specific sites. As discussed by Nolan and co-workers (unit Unavailable ), one of the unique features of flow cytometry is its ability to perform correlated measurements of multiple phosphorylation states at the single cell level. This provides insight into the complexity of signaling networks that is not obtained by standard biochemical techniques. Furthermore, in combination with other phenotypic markers, flow cytometry can measure alterations in signaling pathways in subpopulations of cells. This clearly has wide potential for studying disorders of the hematopoietic and immune systems. Curr. Protocol. Cytom. 46:9.27.1-9.27.19. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • whole blood fixation/permeabilization;
  • signal transduction;
  • phospho-specific antibody;
  • pharmacodynamics;
  • human leukemias