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Titlebook: Receptor Signal Transduction Protocols; Gary B. Willars,R. A. John Challiss Book 20042nd edition Humana Press 2004

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Identification of G-Protein-Coupled Receptor mRNA Expression by Northern Blotting and , Hybridizatis the ability to identify genetic splice variants, or to study multiple RNA molecules sequentially in the same tissue samples..These protocols give step-by-step instructions for the performance of these techniques, and the analysis of the data that can be obtained using them.
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Generation of Model Cell Lines Expressing Recombinant G-Protein-Coupled Receptors,ran, cationic lipids, and electroporation), allowing their transient or stable expression in diverse cell lines. This chapter gives a brief overview of the different techniques and provides methodology for the generation of transiently transfected cells and for selection, isolation and maintenance of stable transfected cell lines.
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Radioligand-Binding Methods for Membrane Preparations and Intact Cells,ing assays for G-protein-coupled receptors in membranes and intact cells are presented for these types of experiments. Detailed procedures for analysis of the data obtained from these experiments are also given.
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Radioligand-Binding Methods for Membrane Preparations and Intact Cells,igand-binding experiments: (1) saturation experiments from which the affinity of the radioligand for the receptor and the binding site density can be determined; (2) inhibition experiments from which the affinity of a competing, unlabeled compound for the receptor can be determined; and (3) kinetic
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