With its detailed description of membrane protein expression, high-throughput and genomic-scale expression studies, both on the analytical and the preparative scale, this book covers the latest advances in the field. The step-by-step protocols and practical examples given for each method constitute practical advice for beginners and experts alike.
This new edition provides a guide to producing crystals of proteins and nucleic acids of sufficient quantity and quality for diffraction studies. It has been thoroughly updated to include all new techniques such as uses of molecular biology in structural biology (sequence modifications to enable crystallization), diagnostic analysis of prenucleation and nucleation by spectroscopic methods, and the two-dimensional electron crystallography of soluble proteins on planar lipid films.
This is an introduction to proteomics, a hot area in the biological sciences which enables scientists to study the protein complement of the genome. It refers to studying the functions of many proteins at one time, as opposed to how research was conducted in the mid-90′s where the technology did not enable the study of the entire protein system. From this, one is able to develop an overview of the way a genome functions and the role it plays in health and disease.
This volume describes reproducible methods for detecting and analyzing the posttranslational modifications of protein, particularly with regard to protein function, proteome research, and the characterization of pharmaceutical proteins. Methods include those for analyzing the assignment of disulfide bond sites in proteins, protein N-glycosylation and protein O-glycosylation, and oligosaccharides present at specific single glycosylation sites in a protein. Additional techniques facilitate the analysis of glycosylphosphatidylinositols, lipid modifications, protein phosphorylation and sulfation, protein methylation and acetylation, a-amidation and lysine hydroxylation
Written for those who have some familiarity with separation of water-soluble proteins, but who may not be aware of the problems involved in the isolation of membrane proteins. This book is an essential manual for investigations of structure and function of native membrane proteins.
Details of the expression of cloned DNA or RNA templates in all the major in vivo and in vitro systems are included such as cultured mammalian cells, the yeasts Saccharomyces cerevisiae and Pichia pastoris, baculovirus, Xenopus oocytes and prokaryotic cells. Cell-free systems of both eukaryotes and prokaryotes are described in addition to the prokaryotic systems that offer coupled transcription-translation.
Here is instant access to over 5000 tested, state-of-the-art protein methods and supporting protocols that are reproducible and accessible on a CD-ROM. Unprecedented in scope and depth of coverage, this collection contains the major procedures needed for isolating, characterizing, and analyzing proteins, including powerful techniques in mass spectroscopy, NMR, proteomics, and immunochemistry. This CD-ROM includes cutting-edge NMR techniques, physical methods of analysis, and protocols for peptide synthesis, membrane proteins, ELISA, proteolytic enzymes, and epitope mapping.
Experts update this classic protocols manual for protein purification techniques. New chapters include those on protein fractionation and chromatographic techniques. New developments in proteomics are included such as the use of 2-D gel electrophoresis as a preparative technique for protein characterization.
They also include newer techniques using analytical chromatography for the multidimensional separations of proteins and peptides as well as mass spectrometric techniques for isolating proteins. The authors provide descriptions of how to scale-up purification methods and the purification of proteins for therapeutic use.
Protein separation techniques have been refined to minimize variability, optimize particular applications, and adapt to user preferences in the analysis of proteins. This book provides an examination of all major separation techniques for proteomics research. A compilation of hands-on methods, it may serve as a guide for selection of the optimal separation strategies to solve particular biological problems.
Progress in the development of robust analytical techniques and instrumentation has created the need for good quality biological samples that are subject to analysis. Emphasizing the importance of sample preparation, the book explains how proteomes can be divided into smaller, less complicated "subproteomes" for individual analysis.