Library Science

A collection of biochemical, cellular, and molecular techniques for unraveling and quantifying the events occurring between the initial contact of a cytokine at the membrane receptor and the eventual activation of gene transcription. The techniques used include the generation of transfectants, the immunohistochemical detection of cytokines in tissue sections, and optimized staining for cytoplasmic detection. Highlights include RT-PCR of small amounts of mRNA, in situ hybridization, biosensor analysis, measurement of biological activities and standardization, immunohistochemical and single-cell detection, and receptor isolation, characterization, and crystallization.

Ed. De Ley, Marc

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An outstanding collection of both classic and cutting-edge techniques for the detection and measurement of mitogen-activated protein kinase (MAPK) signaling cascades. The protocols includes methods for the determination of the subcellular localization of these components, the structural and biophysical analysis of the components, and identification of novel components of known and unknown signaling cascades. Additional methods examine the upstream mechanisms of activation of MAPK cascades by various receptors, the mechanisms involved in the down regulation of the MAPK cascades, and identification of targets of the MAPK cascades. A number of techniques use inhibitors in research on MAPK and transgenic mice for studies of MAPK signaling. The techniques offer biochemists, cell biologists, physicians, and biotechnologists all the essential laboratory techniques needed in to conduct productive studies of MAPK signaling in health and disease, in measuring the influence of drugs, and in a broad range of experimental systems.

Ed. Seger, Rony

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Hands-on experts from academia and industry comprehensively describe how to successfully perform all the critical HPLC techniques needed for the analysis of peptides and proteins. The methods range from commonly used techniques to those for capillary to large-scale preparative isolation. The authors have also presented a number of specific applications as case studies to illustrate the analytical approaches to a particular separation or assay challenge, with examples drawn from contemporary fields in biochemistry and biotechnology.

Ed. Aguilar, Marie-Isabel

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In this completely updated and expanded edition of a classic bench manual, hands-on experts take advantage of the latest advances in ribozyme, DNAzyme, and RNA interference technologies to describe in detail the exciting and successful methods now available for gene inactivation in vitro and in vivo. Their optimized techniques employ hairpain ribozymes, DNAzymes, hammerhead ribozymes and derivatives, group I intron ribozymes, Rnase P ribozymes, and siRNAs, as well as general methods for RNA structure analysis, delivery of oligonucleotides, and gene therapy. Also provided are novel methods for identifying accessible cellular mRNA sites; group I intron and RNAse P ribozymes protocols for effective design, selection, and therapeutic applications; and the latest RNAi methods for sequencing-specific gene silencing in a wide variety of organisms. Comprehensive and up-to-date, Ribozymes and siRNA Protocols synthesizes for experienced and novice investigators alike the exciting advances in understanding nucleic acid enzymes and demonstrates how they may be used to analyze gene function and target validation, and to productively develop new therapeutics for human diseases.

Ed. Sioud, Mouldy

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For volume 1 alone: A gold-standard collection of readily reproducible techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. Volume 1: Sperm and Oocyte Analysis focuses on sperm cells, oocyte analysis, oocyte maturation, fertilization, and preparation techniques. Highlights include in vitro maturation and fertilization of human, porcine, and canine oocytes; the cryopreservation of sperm cells; establishment of an in vitro spermatogenesis system; visualization of sperm accessory structures; and motility assays of stallion spermatozoa. Each readily reproducible protocol is described in step-by-step detail and contains an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. For both volumes: A gold-standard collection of readily reproducible techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. The methods cover sperm and egg activation, motility, fertilization, nuclear development, nuclear cloning, the molecular characterization of specific events, and the imaging of cell structures. Volume 1: Sperm and Oocyte Analysis focuses on sperm cells, oocyte analysis, oocyte maturation, fertilization, and preparation techniques. Volume 2: Molecular Embryo Analysis, Live Imaging, Transgenesis, and Cloning contains methods for molecular egg analysis, live egg imaging, nuclear cloning, oocyte preservation, and nuclear transfer. Comprehensive and cutting-edge, Germ Cell Protocols offers both novice and established researchers a gold-standard collection of hard-to-find methods of high impact research, diverse procedures that are easy-to-follow, well-illustrated, and allow a cross-species transfer of knowledge from lower vertebrates to higher mammalian systems.

Ed. Schatten, Heide

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For volume 2 alone: A gold-standard collection of readily reproducible techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. Volume 2: Molecular Embryo Analysis, Live Imaging, Transgenesis, and Cloning focuses on molecular egg analysis, live egg imaging, nuclear cloning, oocyte cryopreservation, and nuclear transfer. Highlights include the identification and characterization of Oct-4, germline chromatin silencing by RNAi, cDNA subtraction and cloning in the field of trophoblast/placental development, and selective ablation. Each readily reproducible protocol is described in step-by-step detail and contains an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Germ Cell Protocols offers both novice and established researchers a gold-standard collection of hard-to-find methods of high impact research, diverse procedures that are easy-to-follow, well-illustrated, and allow a cross-species transfer of knowledge from lower vertebrates to higher mammalian systems. For both volumes: A gold-standard collection of readily reproducible techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. The methods cover sperm and egg activation, motility, fertilization, nuclear development, nuclear cloning, the molecular characterization of specific events, and the imaging of cell structures. Volume 1: Sperm and Oocyte Analysis focuses on sperm cells, oocyte analysis, oocyte maturation, fertilization, and preparation techniques. Volume 2: Molecular Embryo Analysis, Live Imaging, Transgenesis, and Cloning contains methods for molecular egg analysis, live egg imaging, nuclear cloning, oocyte preservation, and nuclear transfer. Comprehensive and cutting-edge, Germ Cell Protocols offers both novice and established researchers a gold-standard collection of hard-to-find methods of high impact research, diverse procedures that are easy-to-follow, well-illustrated, and allow a cross-species transfer of knowledge from lower vertebrates to higher mammalian systems.

Authors: Schatten, Heide

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