Radioligand binding methods practical guide and tips

D. B. Bylund, Myron Lee Toews

Research output: Contribution to journalArticle

Abstract

Radioligand binding assays are a relatively simple but extremely powerful tool for studying receptors. They allow an analysis of the interactions of hormones, neurotransmitters, growth factors, and related drugs with the receptors, studies of receptor interactions with second messenger systems, and characterization of regulatory changes in receptor number, subcellular distribution, and physiological function. As a result, these assays are widely used (and often misused) by investigators in a variety of disciplines, including pharmacology, physiology, biochemistry, immunology, and cell biology. This article presents a broad overview of the radioligand binding assay technique, primarily for the investigator who has limited experience with this technique. Practical guidelines for setting up a new assay are presented, including the receptor preparation to be used choice of appropriate radioligand, optimizing assay conditions, and appropriate methods for data analysis. Tips for avoiding some of the common pitfalls in application of these assays are also included. The primary focus is on radioligand binding assays of membrane-bound receptors studied in membrane preparations. However, similar assay techniques can be used to study receptors on intact cells. The unique advantages and disadvantages of these intact cell binding assays are also discussed. In particular, the occurrence of regulatory changes in receptors during the course of intact cell binding assays is considered, with approaches for circumventing these complications and for using intact cell assays to advantage in studying these regulatory changes.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume265
Issue number5 9-5
StatePublished - 1993

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Radioligand Assay
Research Personnel
Drug Receptors
Membranes
Second Messenger Systems
Allergy and Immunology
Biochemistry
Neurotransmitter Agents
Cell Biology
Intercellular Signaling Peptides and Proteins
Hormones
Pharmacology
Guidelines

Keywords

  • membrane
  • receptor binding assays
  • receptors

ASJC Scopus subject areas

  • Cell Biology
  • Physiology
  • Pulmonary and Respiratory Medicine

Cite this

Radioligand binding methods practical guide and tips. / Bylund, D. B.; Toews, Myron Lee.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 265, No. 5 9-5, 1993.

Research output: Contribution to journalArticle

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