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Cell Migration
A collection of classic, novel, and state-of-the-art methods for the study of cell migration in cultured cells, different model organisms, and specialized cells in normal development and disease. Highlights include basic assays that apply to all cell migration studies in vitro, assays in various model organisms, and assays for cancer cells, endothelial cells, and neurons both in vitro and in animal models. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Forensic DNA Typing Protocols
A state-of-the-art collection of readily reproducible laboratory methods for DNA identity analysis, including Y chromosome haplotyping, mtDNA, and SNP typing. The book offers well-tested protocols for DNA quantification using real-time PCR on forensic samples and for the determination of the number of amelogenine gene copies. For forensic geneticists, there are readily reproducible methods for species identification, ancient DNA, and pharmacogenetics. Additional chapters address new applications in the forensic genetics lab, such a species identification or typing of CYP polymorphisms for the analysis of adverse to drugs.
Cell Cycle Control
The fundamental question of how cells grow and divide has perplexed biologists since the development of the cell theory in the mid-19th century, when it was recognized by Virchow and others that “all cells come from cells.” In recent years, considerable effort has been applied to the identification of the basic molecules and mechanisms that regulate the cell cycle in a number of different organisms. Such studies have led to the elucidation of the central paradigms that underpin eukaryotic cell cycle control, for which Lee Hartwell, Tim Hunt, and Paul Nurse were jointly awarded the Nobel Prize for Medicine and Physiology in 2001 in recognition of their seminal contributions to this field. The importance of understanding the fundamental mechanisms that modulate cell division has been reiterated by relatively recent discoveries of links between cell cycle control and DNA repair, growth, cellular metabolism, development, and cell death. This new phase of integrated cell cycle research provides further challenges and opportunities to the biological and medical worlds in applying these basic concepts to understanding the etiology of cancer and other proliferative diseases.
Mast Cells
A cutting-edge collection of readily reproducible techniques for the isolation, culture, and study of activation and signaling in human mast cells. These methods take advantage of the latest advances in molecular biology, technology, and information science. They include methods for the identification of mast cells, the development of mast cells in vitro, the study of mast cell signaling and gene expression, and the measurement of mast cell expression of inflammatory mediators. Additional chapters cover methods for studying mast cell interactions with other cell types (endothelial cells, fibroblasts, and B cells), the roles of mast cells in host defense, and mast cell apoptosis.
Peptide Synthesis and Applications
Hands-on experts describe in step-by-step detail the key methodologies of contemporary peptide synthesis and illustrate their numerous applications. The techniques presented include protocols for chemical ligation, the synthesis of cyclic and phosphotyrosine-containing peptides, lipoamino acid- and sugar-conjugated peptides, and peptide purification and analyses. Additional chapters detail methodologies and instrumentation for high-throughput peptide synthesis, many different applications of peptides as novel research tools and biological probes, and the design and application of fluorescent substrate-based peptides that can be used to determine the selectivity and activity of peptidases. A practical guide to the identification of proteins using mass spectrometric analyses of peptide mixtures is also included.
Molecular Toxicology Protocols
A collection of cutting-edge techniques for analyzing genotoxic exposure and detecting the resulting biological effects-including endogenous metabolites-up to and including the development of cancer. The authors emphasize analytical methods that can be specifically applied to human populations and patients. Among the applications detailed are the analysis of interactions between such cellular macromolecules as DNA and proteins and chemical and physical agents, the assessment of medically relevant toxicity, and the characterization of genetic alterations induced in transgenic animals by in vivo systems. There are also methods for the analysis of genotoxic exposure during gene expression, of cytotoxicity caused by the induction of apoptosis, of genetic alterations in reporter genes and oncogenes, early (premalignant) detection of altered oncogenes, and of individual variation in biotransformation and DNA repair capacity.
Chemical Genomics
Chemical genomics is an exciting new field that aims to transform biolo- cal chemistry into a high-throughput industrialized process, much in the same way that molecular biology has been transformed by genomics. The inter- tion of small organic molecules with biological systems (mostly proteins) underpins drug discovery in the pharmaceutical and biotechnology industries, and therefore a volume of laboratory protocols that covers the key aspects of chemical genomics would be of use to biologists and chemists in these orga- zations. Academic scientists have been exploring the functions of proteins using small molecules as probes for many years and therefore would also b- efit from sharing idea...
DNA Repair Protocols
The first edition of this book, published in 1999 and called DNA Repair Protocols: Eukaryotic Systems, brought together laboratory-based methods for studying DNA damage and repair in diverse eukaryotes: namely, two kinds of yeast, a nematode, a fruit fly, a toad, three different plants, and human and murine cells. This second edition of DNA Repair Protocols covers mammalian cells only and hence its new subtitle, Mammalian Systems. There are two reasons for this fresh emphasis, both of them pragmatic: to cater to the interests of what is now a largely mammalocentric DNA repair field, and to expedite editing and prod- tion of this volume. Although DNA Repair Protocols: Mammalian Systems is a s...
Receptor Binding Techniques
A comprehensive collection of readily reproducible methods for studying receptors in silico, in vitro, and in vivo. These cutting-edge techniques cover mining from curated databases, identifying novel receptors by high throughput screening, molecular methods to identify mRNA encoding receptors, radioligand binding assays and their analysis, quantitative autoradiography, and imaging receptors by positron emission tomography (PET). Highlights include phenotypic characterization of receptors in knockout mice, imaging receptors using green fluorescent protein and fluorescent resonance energy transfer, and quantitative analysis of receptor mRNA by TaqMan PCR. These book equips the researcher with techniques for exploring the unprecedented number of new receptor systems now emerging and the so-called "orphan" receptors whose activating ligand has not been identified.
Protein Nanotechnology
Leading experts in nanobiotechnology comprehensively review the most recent advances in instrumentation and methodology, as well as their applications in genomics and proteomics. The authors provide a wide variety of techniques and methods for dealing with protein functions and structures at the nanoscale level, including nanostructured systems, nanomaterials, carbon nanotubes and nanowires, optical nanosensors, and nanoelectrodes. Among the highlights are techniques for the in vivo tracking of biochemical processes using fluorescent molecular probes and nanosensors, and the exploration of biochemical processes and submicroscopic structures of living cells at unprecedented resolutions using near-field optics. Also discussed is the development of nanocarrier methodology for the targeted delivery of drugs whose shells are conjugated with antibodies for targeting specific antigens.
