Adrenergic Receptor ProtocolsCurtis A. Machida Springer Science & Business Media, 3 feb 2008 - 577 páginas Adrenergic receptors are important modulators in the sympathetic c- trol of various metabolic processes in the central and peripheral nervous s- tems. These receptors are localized at multiple sites throughout the central nervous system (CNS) and serve as important regulators of CNS-mediated behavior and neural functions, including mood, memory, neuroendocrine c- trol, and stimulation of autonomic function. Adrenergic Receptor Protocols consists of 35 chapters dealing with va- ous aspects of adrenergic receptor analyses, including the use of genetic, RNA, protein expression, transactivator, second messenger, immunocytochemical, electrophysiological, transgenic, and in situ hybridization approaches. This volume details the use of various methods to examine the adrenergic receptor system, using aspects of the genetic flow of information as a guide (DNA? RNA ? transactivator ? protein expression ? second messenger analyses ? cellular analyses ? transgenic whole animal approaches). Adrenergic Receptor Protocols displays step-by-step methods for s- cessful replication of experimental procedures, and would be useful for both experienced investigators and newcomers in the field, including those beg- ning graduate study or undergoing postdoctoral training. The Notes section contained in each chapter provides valuable troubleshooting guides to help develop working protocols for your laboratory. With Adrenergic Receptor Protocols, it has been my intent to develop a comprehensive collection of modern molecular methods for analyzing adrenergic receptors. I would like to thank the many chapter authors for their contributions. |
Índice
11 | |
Isolation of Adrenergic Receptor Genes | 37 |
Analyses of Adrenergic Receptor Sequences | 53 |
Polymerase Chain Reaction Screening of Genomic Libraries | 73 |
SolutionPhase Library Screening for Identification | 91 |
Genetic Polymorphisms of Adrenergic Receptors | 117 |
Northern Blot Analyses Detecting Adrenergic Receptor mRNAs | 129 |
Ribonuclease Receptor | 139 |
Transient Transfection and Adrenergic Receptor Promoter Analysis | 235 |
Targeted Disruption of Adrenergic Receptor Genes | 259 |
Development of Antibodies to Adrenergic Receptors | 281 |
Dunigan Patricia K Curran and Peter H Fishman | 329 |
Assessing Adrenergic Receptor Conformation Using Chemically | 345 |
Anne Dam Jensen and Ulrik Gether 24 Biochemical Methods for Detection and Measurement | 363 |
Assay of Arachidonic Acid Release Coupled | 375 |
Lincoln Edwards and Paul Ernsberger 26 PatchClamp Recording Methods for Examining Adrenergic | 391 |
Determination of Adrenergic Receptor mRNAs by Quantitative | 155 |
Nuclear RunOn Assays for Measurement of Adrenergic Receptor | 169 |
for Determination of | 181 |
Use of Eukaryotic Vectors for the Expression of Adrenergic Receptors | 189 |
Expression of βAdrenergic Receptors in Recombinant | 207 |
Expression of βAdrenergic Receptors in E coli | 215 |
Use and Pharmacological Analysis of Established | 221 |
Southwestern Blots for Detection of a DNA Binding Protein | 411 |
Recognizing the | 419 |
Philbert Kirigiti and Curtis A Machida 30 Determination of mRNA Stability and Characterization of Proteins | 453 |
Use of Immunohistochemistry and Confocal Microscopy | 469 |
In Situ Hybridization of Adrenergic Receptor mRNA in Brain | 519 |
Use of Electron Microscopy in the Detection of Adrenergic Receptors | 535 |
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acetate adrenergic receptor agarose agonist alignment aliquots amino acid amplified analysis antibody antisense RNA assay binding Biol blot buffer cDNA cell lines centrifuge Chem cloning concentration containing culture denaturing detection determined digestion dilution DNA polymerase EDTA electrophoresis enzyme ethanol expression extract filter fragments G-protein-coupled receptors genomic DNA human hybridization Incubate insert labeling ligation medium membrane Methods in Molecular mg/mL Molecular Biology mRNA NaCl nitrocellulose Note nucleotide ODNs oligonucleotide PCR product pellet peptide phage Pharmacol phenol photoaffinity labels pipet plasmid plate polymerase polymorphisms prepared primer probe protein Protocols purified radioligand reaction reagents receptor subtypes recombinant restriction enzyme Resuspend RNase room temperature sample screening sequence serum sodium Southern blot specific sterile stored Subheading supernatant TAE buffer target template tion tissue transcription transfection transfer Tris-HCl tube vector volume wash µg/mL