Unit

UNIT 12.4 Cellular Assays of Chemokine Receptor Activation

  1. Amanda E.I. Proudfoot1,
  2. Christine A. Power1,
  3. Dennis J. Church1,
  4. Dulce Soler2,
  5. Matthias Mack3

Published Online: 1 NOV 2001

DOI: 10.1002/0471141755.ph1204s14

Current Protocols in Pharmacology

Current Protocols in Pharmacology

How to Cite

Proudfoot, A. E., Power, C. A., Church, D. J., Soler, D. and Mack, M. 2001. Cellular Assays of Chemokine Receptor Activation. Current Protocols in Pharmacology. 14:12.4:12.4.1–12.4.26.

Author Information

  1. 1

    Serono Pharmaceutical Research Institute, Plan-les-Ouates, Switzerland

  2. 2

    Millennium Pharmaceuticals, Cambridge, Massachusetts

  3. 3

    Ludwig-Maximilians-University of Munich, Munich, Germany

Publication History

  1. Published Online: 1 NOV 2001
  2. Published Print: SEP 2001

Abstract

This unit describes procedures for measuring the activation of chemokine receptors by their ligands. Chemokines are chemoattractant proteins and two assays are described for analyzing the chemoattractant properties (chemotaxis) of these proteins. Chemokine receptors are coupled to G proteins, and activation of the receptors results in the mobilization of intracellular Ca2+ stores. Two assays for the measurement of this activity are described which employ calcium-sensitive dyes, one for adherent and one for non-adherent cells. In addition, an assay for quantitating chemokine receptor downmodulation and subsequent recycling events is described. The assays are applicable to both purified leukocytes and stably expressed recombinant chemokine receptors.This unit describes procedures for measuring the activation of chemokine receptors by their ligands.