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.
Includes supplementary material: sn.pub/extras
Klappentext
Maintaining the high standards set by the successful first edition, Anthony P. Davenport and a panel of hands-on researchers from the pharmaceutical industry and academia take advantage of the latest developments to provide detailed practical methods for studying receptors in silico, in vitro, and in vivo. These readily reproducible 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 knock-out mice, imaging receptors using green fluorescent protein and fluorescent resonance energy transfer, and quantitative analysis of receptor mRNA by TaqMan polymerase chain reaction (PCR). These ligand binding techniques are perfect for exploring the unprecedented number of new receptor systems now emerging and the so-called "orphan" receptors whose activating ligand has not been identified. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and state of the art, Receptor Binding Techniques, Second Edition offers academic and commercial researchers in the pharmaceutical and biotechnology industries a set of proven techniques for the successful characterization of receptors and the phenotyping of transgenic animals, including knock-outs.
Inhalt
Receptor Databases and Computational Websites for Ligand Binding Brinda K. Rana and Paul A. Insel Identification of Orphan G Protein-Coupled Receptor Ligands Using FLIPR® Assays Nicola M. Robas and Mark D. Fidock Quantitative Analysis of Orphan G Protein-Coupled Receptor mRNAs by TaqMan® 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 mRNA: Detection by In Situ and Northern Hybridization Alessandra P. Princivalle, Rachel M. C. Parker, Terri J. Dover, and Nicholas M. Barnes Radioligand-Binding and Molecular-Imaging Techniques for the Quantitative Analysis of Established and Emerging Orphan Receptor Systems Anthony P. Davenport and Rhoda E. Kuc Measurement of Radioligand Binding by Scintillation Proximity Assay Jenny Berry and Molly Price-Jones Autoradiography of Enzymes, Second Messenger Systems, and Ion Channels David A. Walsh and John Wharton 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 Live Cell Imaging of G Protein-Coupled Receptors Burkhard Wiesner, Michael Beyermann, and Alexander Oksche Imaging and Characterization of Radioligands for Positron Emission Tomography Using Quantitative Phosphor Imaging Autoradiography Peter Johnström and Anthony P. Davenport Dynamic In Vivo Imaging of Receptors in Small Animals Using Positron Emission Tomography Peter Johnström, Tim D. Fryer, Hugh K. Richards, Olivier Barret, and Anthony P. Davenport Index