This fully updated and expanded second edition of Transmembrane Signaling Protocols demonstrates the use of such techniques as single cell imaging, bioluminecence resonance energy transfer, and global proteomics in the study of transmembrane signaling events. Highlights include the functional expression and genetic selection of mutant mammalian transmembrane receptors in yeast, real-time analysis of transmembrane receptor signaling in live cells, and bioluminescence energy transfer to monitor protein-protein interaction. Additional chapters use state-of-the-art methods to analyze gene expression and proteomic profiles related to transmembrane signaling events, as well as to genetically reconstitute bone marrow for the study of signal transduction ex vivo. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.The complete sequencing of the human genome and many new microarray and proteomic methods for analyzing global changes in both gene expression and protein profiles have greatly increased our ability to understand transmembrane signaling processes in close detail. In this fully updated and expanded second edition of Transmembrane Signaling Protocols, leading experts demonstrate the use of such techniques as single-cell imaging, bioluminescence resonance energy transfer, and global proteomics in the study of transmembrane signaling events. Highlights include the functional expression and genetic selection of mutant mammalian transmembrane receptors in yeast, real-time analysis of transmembrane receptor signaling in live cells, and bioluminescence energy transfer to monitor protein-protein interaction. Additional chapters use state-of-the-art methods to analyze gene expression and proteomic profiles related to transmembrane signaling events, as well as to genetically reconstitute bone marrow for the study of signal transduction ex vivo. Introductory chapters provide a wide-ranging perspective, including an overview of the many recent advances in our understanding of signal transduction pathways. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. §Cutting-edge and highly practical, Transmembrane Signaling Protocols, Second Edition, makes available to novice and experienced researchers alike the readily reproducible techniques needed to advance the study of signal transduction pathways today.
