Receptor Signal Transduction Protocols

Inducible Expression of G Protein-Coupled Receptors in Transfected Cells.- Using the Flp-InTM T-RexTM System to Regulate GPCR Expression.- Viral Infection for GPCR Expression in Eukaryotic Cells.- Generation of Epitope-Tagged GPCRs.- The Use of Site-Directed Mutagenesis to Study GPCRs.- Approaches to Study GPCR Regulation in Native Systems.- Heterologous Expression of GPCRs in Fission Yeast.- Radioligand Binding Methods for Membrane Preparations and Intact Cells.- Quantification of GPCR mRNA Using Real-Time RT-PCR.- Determining Allosteric Modulator Mechanism of Action: Integration of Radioligand Binding and Functional Assay Data.- Design and Use of Fluorescent Ligands to Study Ligand-Receptor Interactions in Single Living Cells.- Examining Site-Specific GPCR Phosphorylation.- Ubiquitination of GPCRs.- [35S]GTPGammaS Binding as an Index of Total G-Protein and G -Subtype-Specific Activation by GPCRs.- Using Calcium Imaging as a Readout of GPCR Activation.- Measuring Spatio-Temporal Dynamics of Cyclic AMP Signaling in Real-Time Using FRET-Based Biosensors.- Determining the Activation of Rho as an Index of Receptor Coupling to G12/13 Proteins.- The Use of Translocating Fluorescent Biosensors for Real-Time Monitoring of GPCR-Mediated Signaling Events.- Study of GPCR-Protein Interactions Using Gel Overlay Assays and Glutathione-S-Transferase-Fusion Protein Pull-Downs.- Study of GPCR-Protein Interactions by BRET.- Time Resolved FRET Strategy with Fluorescent Ligands to Analyze Receptor Interactions in Native Tissues: Application to GPCR Oligomerization.- Peptide Affinity Purification for the Isolation and Identification of GPCR-AssociatedProtein Complexes.- Tandem Affinity Purification and Identification of GPCR-Associated Protein Complexes.- Identification of GPCR Localization in Detergent Resistant Membranes.- Analysis of GPCR Localization and Trafficking.- Statistical Methods in Research.

From the reviews of the third edition:"This is an extensive collection of established, new, and updated techniques for the study of the consequences of receptor activation. Each of the 26 chapters offers a specific recipe, with lists of required reagents and equipment, plus notes to aid in use of the protocol. ... This book would be most useful to researchers already familiar with fundamental molecular biological or biochemical laboratory techniques." (Kenneth E. McCarson, Doody's Book Reviews, February, 2012)