Receptor Binding Techniques (Methods in Molecular Biology)

A broad definition of a receptor is a specialized protein on or in a cell that recognizes and binds a specific ligand  to undergo a conformational change, leading to a physiological response or change in cell function.  A ligand can be an endogenous neurotransmitter, hormone, paracrine/autocrine factor, or a synthetic drug that may function as an agonist or antagonist. The third edition of  Receptor Binding Techniques expands upon the methods and techniques used for studying receptors in silico, in vitro and in vivo. Comprehensive chapters describe how to use online resources for experimental research such as prediction of receptor-ligand interactions and mine the IUPHAR receptor database. Classical techniques of radioligand binding, quantitative autoradiography and their analyses are complemented by the use of  immunocytochemistry  for the cellular localization of receptor protein and hybridization to detect receptor mRNA.  Protocols using fluorescent labeled ligands are described to visualise receptors in living cells, their interaction with beta-arrestin to measure ligand-induced internalisation and  green fluorescent protein to study  trafficking. Non-radioactive, chemiluminescent cAMP and arrestin assays facilitate the identification of novel ‘biased agonists’. Detailed methods are provided for in vivo imaging of receptors using positron emission tomography (PET). Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Receptor Binding Techniques, Third Edition, aids scientists in continuing to study receptor binding.

Receptor Binding Techniques (Methods in Molecular Biology)

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Receptor Binding Techniques (Methods in Molecular Biology (Cloth))

This cutting-edge collection of step-by-step experimental protocols demonstrates the power of radioligand binding for studying a wide variety of receptors. The methods can be applied to G protein coupled receptors, receptors with integral ion channels, and sites other than receptors, such as guanine nucleotide binding proteins, enzymes, transporters, and mRNA. The individual protocols encompass many different preparations, ranging from solubilized receptors, through membrane preparations, intact cells, and autoradiography in tissue slices, to PET and SPECT analysis of binding in living human brain. Along with the detailed, easily reproducible instructions, the protocols also address the problems inherent in the basic binding technique, such as common experimental artifacts and the definition of receptor sites, as well as problems associated with the use of particular ligands and receptor preparations. Receptor Binding Techniques shows clearly how these methods can be adapted to many different applications and fully utilized in drug discovery programs.

Receptor Binding Techniques (Methods in Molecular Biology (Cloth))

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Receptor Binding Techniques (Methods in Molecular Biology)

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Receptor Binding Techniques (Methods in Molecular Biology) Overview

A comprehensive collection of readily reproducible methods for studying receptors in silico, in vitro, and in vivo. These cutting-edge techniques cover mining from curated databases, identifying novel receptors by high throughput screening, molecular methods to identify mRNA encoding receptors, radioligand binding assays and their analysis, quantitative autoradiography, and imaging receptors by positron emission tomography (PET). Highlights include phenotypic characterization of receptors in knockout mice, imaging receptors using green fluorescent protein and fluorescent resonance energy transfer, and quantitative analysis of receptor mRNA by TaqMan PCR. These book equips the researcher with techniques for exploring the unprecedented number of new receptor systems now emerging and the so-called "orphan" receptors whose activating ligand has not been identified.

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