About this title: 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.
Table Of Contents
Preface
Contributors
1 Receptor Databases and Computational Websites for Ligand Binding
Brinda K. Rana and Paul A. Insel
2 Identification of Orphan G Protein-Coupled Receptor Ligands Using FLIPR?Assays
Nicola M. Robas and Mark D. Fidock
3 Quantitative Analysis of Orphan G Protein-Coupled Receptor mRNAs by TagMan?Real-Time PCR: G2A and GPR4 Lysophospholipid Receptor Expression in Leukocytes and in a Rat Myocardial Infarction-Heart Failure Model
Stephen A. Douglas, Zhaohui Ao, Douglas G. Johns, Kristeen Maniscalco, Robert N. Willette, Lea Sarov-Blat, John P. Cogswell, Sheila Seepersaud, Paul Murdock, Klaudia M. Steplewski, and Lisa Patel
4 mRNA: Detection by In Situ and Northern Hybridization
Alessandra P. Princivalle, Rachel M.C. Parker, Terri J. Dover, and Nicholas M. Barnes
5 Radioligand-Binding and Molecular-Imaging Techniques for the Quantitative Analysis of Established and Emerging Orphan Receptor Systems
Anthony P. Davenport and Rhoda E. Kuc
6 Measurement of Radioligand Binding by Scintillation Proximity Assay
Jenny Berry and Molly Price-Jones
7 Autoradiography of Enzymes, Second Messenger Systems, and Ion Channels
David A. Walsh and John Wharton
8 Immunocytochemical Localization of Receptors Using Light and Confocal Microscopy With Application to the Phenotypic Characterization of Knock-Out Mice
Anthony P. Davenport and Rhoda E. Kuc
9 Live Cell Imaging of G Protein-Coupled Receptors
Burkhard Wiesner, Michael Beyermann, and Alexander Oksche
10 Imaging and Characterization of Radioligands for Positron Emission Tomography Using Quantitative Phosphor Imaging Autoradiography
Peter Johnstr? and Anthony P. Davenport
11 Dynamic In Vivo Imaging of Receptors in Small Animals Using Positron Emission Tomography
Peter Johnstr?, Tim D. Fryer, Hugh K. Richards, Olivier Barret, and Anthony P. Davenport
Index
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