Polyamine Protocols


Book Description

A unique collection of hands-on enzyme assay techniques to study polyamines and their function. The techniques range from assay methods for enzymes of polyamine biosynthesis and catabolism to measurements of polyamines, polyamine transport, and polyamine effects on cell growth. The methods are presented by leading researchers who have perfected them to a high art, and include clear, step-by-step instructions with numerous hints and tips to ensure readily reproducible results.




Polyamines in Health and Nutrition


Book Description

The importance of polyamines for all living cells has been recognized since spermine was discovered in human semen more than 300 years ago. Polyamine research intensified when analytical methods were developed for their determination, particularly in tissues and biological fluids. Discovering their close correlation with cancer, and that polyamine concentrations change during the cell cycle, gave reason for further research in this topic. Polyamines in Health and Nutrition concentrates on the direction of polyamine research which has the capacity to influence and benefit our health and which can explain some of the discrepancies and failures of earlier research. It is important to recognize the dietary contribution to the polyamine body pool and to investigate how the polyamine content of the diet can be changed, with the ultimate aim of using this information to improve our health.




Polyamines


Book Description

Recently, important new findings in the polyamine field and a variety of new experimental systems have revolutionized the study of these ubiquitous cellular components, essential for normal growth and development. In Polyamines: Methods and Protocols, leading researchers contribute an extensive collection of up-to-date laboratory techniques for the further pursuit of polyamine study. The volume delves into vital subjects such as neoplasia studies with animal models and human patients, therapeutic roles for polyamine inhibitors and analogs, polyamine metabolism and oxidative damage, polyamines as regulators of critical ion channels, as well as polyamine transport systems and polyamine-responsive genes. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and expert notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Polyamines: Methods and Protocols provides a key resource for all scientists pursuing the study of this dynamic and significant aspect of cellular biology.




The Handbook of Microbial Metabolism of Amino Acids


Book Description

This book collates and reviews recent advances in the microbial metabolism of amino acids, emphasizing diversity - in terms of the range of organisms under investigation and their natural ecology - and the unique features of amino acid metabolism in bacteria, yeasts, fungi, protozoa and nematodes. As well as studying the individual amino acids, including arginine, sulfur amino acids, branched-chain amino acids and aromatic amino acids, a number of themes are explored throughout the work. As the volume of research into the metabolism of amino acids grows, this comprehensive study of the subject is a vital tool for researchers in the fields of biological, medical and veterinary sciences, including microbiology, biochemistry, genetics and pathology. This book is also essential for corporate organizations with active research and development programmes, such as those in the pharmaceutical industry.




2-D Proteome Analysis Protocols


Book Description

With the completion of sequencing projects and the advancement of a- lytical tools for protein identification, proteomics—the study of the expressed part of the genome—has become a major region of the burgeoning field of functional genomics. High-resolution 2-D gels can reveal virtually all p- teins present in a cell or tissue at any given time, including posttranslationally modified proteins. Changes in the expression and structure of most cellular proteins caused by differentiation or external stimuli can be displayed and eventually identified using 2-D protein gels. 2-D Proteome Analysis Protocols covers all aspects of the use of 2-D protein electrophoresis for the analysis of biological problems. The contri- tors include many of the leaders in the fields of biochemistry and analytical chemistry who were instrumental in the development of high-resolution 2-D gels, immobilized pH gradients, computer analysis, and mass spectromet- based protein identification methodologies. This book is intended as a benchtop manual and guide both for novices to 2-D gels and for those aficionados who wish to try the newer techniques. Any group using protein biochemistry—especially in the fields of molecular biology, biochemistry, microbiology, and cell biology—should find this book eminently useful. 2-D Proteome Analysis Protocols takes the researcher through the c- plete process of working with 2-D protein gels from making the protein - tract to finally identifying the proteins of interest. It includes protocols for generating 2-D protein extracts from most of the standard model organisms, including bacteria, yeast, nematode, Drosophila, plants, mouse, and human.




Glycoanalysis Protocols


Book Description

Now fully updated and considerably expanded, Glycoanalysis Protocols, 2nd ed., makes available to all protein scientists, and particularly those working with today's pharmaceuticals, the most advanced and reproducible glycoanalysis techniques currently in use. Developed by highly experienced carbohydrate chemists, biochemists, and physical chemists, these detailed, up-to-date, and proven analytical techniques cover the areas of glycoprotein macromolecular structural analysis, oligosaccharide profiling, lipid conjugate characterization, microorganism structure determination, and proteoglycan function. Special attention has been given to advanced analytical techniques in biotechnology during the production of recombinant glycoproteins and other therapeutics. Hailed as "indispensable" in its first edition, Glycoanalysis Protocols, 2nd ed., continues with vital, time-tested techniques addressing the needs of both biomedical researchers and protein macromolecular structural chemists. It will well serve all those starting work on the analysis of glycoproteins, as well as more experienced investigators seeking to augment their expertise.




Immunochemical Protocols


Book Description

This much anticipated second edition provides a user-friendly, up-to-date handbook of reliable immunochemical techniques optimized for molecular biologists. It covers the breadth of relevant established methods from protein blotting and immunoassays through to visualization of cellular antigens and in situ hybridization, each with their latest refinements. Protocols for the production and purification of important classes of immunochemical reagents are also provided, including "conventional" and recombinant antibodies, fusion proteins and their various conjugates. This book will open the door to a new generation of immunochemical reagents with exciting possibilities.




Nitric Oxide Protocols


Book Description

Michael Titheradge provides both the experienced researcher and the newcomer with time-tested techniques to cover all aspects of NO synthase and metabolism. Topics range from the cloning and expression of the different isoforms of NO synthase to the role of NO in DNA damage and apoptosis and include indirect and direct measurements of NO production, measurement on NO synthase mRNA both in situ and in vitro, and Western blotting and immunohistochemical localization of different isoforms of NO synthase. A variety of alternative techniques are described to enable researchers to set up an assay using either the simplest equipment or the most expensive EPR machines.




Cytochrome P450 Protocols


Book Description

In Cytochrome P450 Protocols, Ian Phillips and Elizabeth Shephard assemble a comprehensive collection of cutting-edge techniques for the investigation of cytochromes P450. Described in detail by hands-on experimentalists for easy reproducibility, these methods include spectral analysis, purification and enzymatic assays, expression in heterologous systems, and the production and use of antibodies, as well as methods for quantification of gene expression, transfection of hepatocytes, and for the investigation of DNA-protein interactions and genetic polymorphisms. In addition, because of the growing importance of in vitro systems in pharmacological toxicology, the book contains techniques for the culture of rodent and human hepatocytes and human epidermis. Cytochrome P450 induction as a biomarker for environmental pollution and the generation of mice with targeted gene disruptions complete this exhaustive collection of core techniques. Cytochrome P450 Protocols includes in one volume both state-of-the-art and classic methods that have not been superseded but remain extremely useful. The collection provides both novice and experienced researchers across many fields-toxicology, pharmacology, environmental biology, biochemistry, and molecular biology-all the tools needed to elucidate the crucial biological role played by cytochromes P450 in the metabolism of therapeutic drugs, chemical carcinogens, and environmental pollutants.




Protein Targeting Protocols


Book Description

It is by no means a revelation that proteins are not uniformly distributed throughout the cell. As a result, the idea that protein molecules, because of the specificity with which they can engage in interactions with other proteins, may be aimed—via these interactions—at a restricted target, is a fundamental one in contemporary molecular life sciences. The target may be variously c- ceived as a specific molecule, a group of molecules, a structure, or a more generic type of intracellular environment. Because the concept of protein targeting is intuitive rather than expl- itly defined, it has been variously used by different groups of researchers in cell biology, biochemistry, and molecular biology. For those working in the field of intracellular signaling, an influential introduction to the topic was the seminal article by Hubbard & Cohen (TIBS [1993] 18, 172–177), which was based on the work of Cohen’s laboratory on protein phosphatases. Sub- quently, the ideas that they discussed have been further developed and extended by many workers to other key intermediaries in intracellular sign- ing, including protein kinases and a great variety of modulator and adaptor proteins.