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Radioligand Binding Methods for Membrane Preparations and Intact Cells

  • David B. BylundEmail author
  • Myron L. Toews
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 746)

Abstract

The radioligand binding assay is a relatively simple but powerful tool for studying G protein-coupled receptors. There are three basic types of radioligand binding experiments: (1) saturation experiments from which the affinity of the radioligand for the receptor and the binding site density can be determined; (2) inhibition experiments from which the affinity of a competing, unlabeled compound for the receptor can be determined; and (3) kinetic experiments from which the forward and reverse rate constants for radioligand binding can be determined. Detailed methods for typical radioligand binding assays for G protein-coupled receptors in membranes and intact cells are presented for these types of experiments. Detailed procedures for analysis of the data obtained from these experiments are also given.

Key words

Affinity Assay Binding Competition G protein-coupled receptor Inhibition Intact cell Kinetic Non-specific binding Radioligand Rate constant Receptor Radioreceptor Saturation 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Pharmacology and Experimental NeuroscienceUniversity of Nebraska Medical CenterOmahaUSA

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