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This second edition of Receptor Signal Transduction Protocols not only has a new editor, but also a greater focus on G-protein-coupled receptors, their properties per se, and their coupling to immediate downstream binding partners—principally, although not exclusively, the heterotrimeric G-proteins. The new edition combines updates of key chapters from the first edition, as well as a large number of new contributions covering key methodologies that have emerged, or been extended to receptor/G-protein research, in the past 5–6 years. In common with many fields, the range of methods used to assess the first steps in signal transduction are continually expanding and methods that might have ...
In the first edition of Calcium Signaling Protocols I began by writing “The regula- 2+ tion of intracellular Ca is a common theme presented in many papers over the last 20 2+ or so years and the description of the Ca -sensitive indicator dye fura-2 in 1985 resulted in a massive increase in these types of studies. ” This statement is as true in 2005 as it was in 1999, but 20 or so years is now 30 years! There has been some reorganization of the volume such that there are now 22 ch- ters including five new ones, all written by experts in their field. These new chapters 2+ include use of the FlexStation and electrophysiological measurement of Ca channel activity. The book is broken into six...
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As the mysteries stored in our DNA have been more completely revealed, scientists have begun to face the extraordinary challenge of unraveling the int- cate network of protein–protein interactions established by that DNA fra- work. It is increasingly clear that proteins continuously interact with one another in a highly regulated fashion to determine cell fate, such as proliferation, diff- entiation, or death. These protein–protein interactions enable and exert str- gent control over DNA replication, RNA transcription, protein translation, macromolecular assembly and degradation, and signal transduction; essentially all cellular functions involve protein–protein interactions. Thus, pro...
Experienced researchers describe in step-by-step detail methods that have proven most useful in delivering genes to mammalian cells. Volume 1 focuses on gene delivery by a variety of chemical and physical methods, including ultrasound, biolistics, peptides, PNA clamps, liposomes, microinjection, electroporation, particle bombardment, dendrimers, and hydrodynamics. Volume 2 details procedures for delivering genes to cells in vitro and in vivo, including the use of lentiviral vectors.
Patch Clamp Methods and Protocols surveys the typical patch clamp applications and advises scientists on identifying problems and selecting the best technique in each instance. The experiments described require a basic level of electrophysiological training and aid the researcher in pursuing new areas of electrophysiology and using the patch clamp technique effectively. Patch Clamp Methods and Protocols is divided into three sections that cover the major areas of patch clamp application: Pharmacology, Physiology, and Biophysics. The first section provides examples and step by step instructions on how to use whole-cell and single-channel patch clamp methods for testing drugs in industrial set...
Flow cytometry has evolved since the 1940s into a multidisciplinary field incorporating aspects of laser technology, fluid dynamics, electronics, optics, computer science, physics, chemistry, biology, and mathematics. Innovations in instrumentation, development of small lasers, discovery of new fluorochromes/fluorescent proteins, and implementation of novel methodologies have all contributed to the recent rapid expansion of flow cytometry applications. In this thoroughly revised and updated second edition of Flow Cytometry Protocols, time-proven as well as cutting-edge methods are clearly and comprehensively presented by leading experimentalists. In addition to being a valuable reference man...
Cells possess a wealth of posttranscriptional control mechanisms that impact on every conceivable aspect of the life of an mRNA. These processes are intimately intertwined in an almost baroque manner, where promoter context influences the recruitment of splicing factors, where the majority of pre-mRNAs undergo alternative splicing, and where proteins deposited during nuclear processing impact distal cytoplasmic processing, translation, and decay. If there is a unifying theme to mRNA Processing and Metabolism: Methods and Protocols, it is that mRNA processing and metabolism are integrated processes. Many of the techniques used to study mRNA have been described in a previous volume of this ser...
Alan V. Smrcka presents a collection of cutting-edge methods for investigating G protein signaling from a variety of perspectives ranging from in vitro biochemistry to whole animal studies. Among the readily reproducible techniques presented are those for the purification of G proteins and effectors enzymes, assays of these purified G proteins and effector enzymes, and for the study of G proteins interactions with effectors in intact cells. Additional methods are provided for assaying G protein coupled receptor structure, function, and localization, and for studying the physiological roles for endogenous G proteins.
Parasitic diseases remain a major health problem throughout the world, for both humans and animals. For many of us, our technologically advanced lifestyle has decreased the prevalence and transmission of parasitic diseases, but for the majority of the world’s population, they are ever present in homes, domestic animals, food, or the environment. The study of parasites and parasitic disease has a long and distinguished history. In some cases, it has been driven by the great importance of the presence of the parasite to the community, for example, those that affect our livestock. In other cases, it is clear that applied research has suffered for lack of funding because the parasite affects p...