Combinatorial Peptide Library Protocols

Combinatorial Peptide Library Protocols

Author: Shmuel Cabilly

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 320

ISBN-13: 1592595715

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During the course of evolution, an imbalance was created between the rate of vertebrate genetic adaptation and that of the lower forms of living organisms, such as bacteria and viruses. This imbalance has given the latter the advantage of generating, relatively quickly, molecules with unexpected structures and features that carry a threat to vertebrates. To compensate for their weakness, vertebrates have accelerated their own evolutionary processes, not at the level of whole organism, but in specialized cells containing the genes that code for antibody molecules or for T-cell receptors. That is, when an immediate requirement for molecules capable of specific interactions arose, nature has preferred to speed up the mode of Darwinian evolution in pref- ence to any other approach (such as the use of X-ray diffraction studies and computergraphic analysis). Recently, Darwinian rules have been adapted for test tube research, and the concept of selecting molecules having particular characteristics from r- dom pools has been realized in the form of various chemical and biological combinatorial libraries. While working with these libraries, we noticed the interesting fact that when combinatorial libraries of oligopeptides were allowed to interact with different selector proteins, only the actual binding sites of these proteins showed binding properties, whereas the rest of the p- tein surface seemed "inert. " This seemingly common feature of protein- having no extra potential binding sites--was probably selected during evolution in order to minimize nonspecific interactions with the surrounding milieu.


Combinatorial Library

Combinatorial Library

Author: Lisa B. English

Publisher: Springer Science & Business Media

Published: 2008-02-04

Total Pages: 380

ISBN-13: 1592592856

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The continued successes of large- and small-scale genome sequencing projects are increasing the number of genomic targets available for drug d- covery at an exponential rate. In addition, a better understanding of molecular mechanisms—such as apoptosis, signal transduction, telomere control of ch- mosomes, cytoskeletal development, modulation of stress-related proteins, and cell surface display of antigens by the major histocompatibility complex m- ecules—has improved the probability of identifying the most promising genomic targets to counteract disease. As a result, developing and optimizing lead candidates for these targets and rapidly moving them into clinical trials is now a critical juncture in pharmaceutical research. Recent advances in com- natorial library synthesis, purification, and analysis techniques are not only increasing the numbers of compounds that can be tested against each specific genomic target, but are also speeding and improving the overall processes of lead discovery and optimization. There are two main approaches to combinatorial library production: p- allel chemical synthesis and split-and-mix chemical synthesis. These approaches can utilize solid- or solution-based synthetic methods, alone or in combination, although the majority of combinatorial library synthesis is still done on solid support. In a parallel synthesis, all the products are assembled separately in their own reaction vessels or microtiter plates. The array of rows and columns enables researchers to organize the building blocks to be c- bined, and provides an easy way to identify compounds in a particular well.


Protein Targeting Protocols

Protein Targeting Protocols

Author: Roger A. Clegg

Publisher: Springer Science & Business Media

Published: 2008-02-04

Total Pages: 331

ISBN-13: 1592595723

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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.


2-D Proteome Analysis Protocols

2-D Proteome Analysis Protocols

Author: Andrew J. Link

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 585

ISBN-13: 1592595847

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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.


Integrated Drug Discovery Technologies

Integrated Drug Discovery Technologies

Author: Houng-Yau Mei

Publisher: CRC Press

Published: 2002-03-19

Total Pages: 603

ISBN-13: 082474473X

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Integrated Drug Discovery Technologies provides a global overview of emerging drug development technologies by presenting and integrating new techniques from the disciplines of chemistry, biology, and computational sciences. It combines integration of contemporary mechanization with strategies in drug delivery. Topics include: functional genomics, microfabrication techniqes, integrated proteomics technologies, high throughput screening, and fluorescence correlation spectroscopy methods.


Polyamine Protocols

Polyamine Protocols

Author: David M. L. Morgan

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 173

ISBN-13: 1592595650

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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.


Transmembrane Signaling Protocols

Transmembrane Signaling Protocols

Author: Dafna Bar-Sagi

Publisher: Springer Science & Business Media

Published: 1998

Total Pages: 631

ISBN-13: 0896034887

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This collection of practical, cutting-edge techniques for the study of cell signaling provides detailed, step-by-step instructions, helpful notes, and troubleshooting tips that make even the most powerful of the newest techniques readily reproducible. The protocols presented include the use of peptide libraries to study transmembrane signaling; the use of single-cell assays to analyze signal transduction pathways; the reconstitution of signaling complexes; methods for analyzing protein-protein interactions, and more. Introductory reviews explain the basic theory and enable researchers new to the area to rapidly gain understanding, as well as command of the practical knowledge and expertise afforded by the protocols. Transmembrane Signaling Protocols makes available to all researchers the many state-of-the-art techniques that have recently led to landmark discoveries in transmembrane signaling.


Forensic DNA Profiling Protocols

Forensic DNA Profiling Protocols

Author: Patrick J. Lincoln

Publisher: Springer Science & Business Media

Published: 1998-01-22

Total Pages: 617

ISBN-13: 0896034437

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This state-of-the-art collection of easily reproducible methods includes all of the major techniques of DNA analysis currently used in forensic identity testing. The methods include the recovery of DNA from a large range of sample types, analysis of DNA as single and multi-locus VNTR probes, PCR amplification of STR and other loci, and mitochondrial sequencing. The expert scientists writing here -- many from laboratories around the world -- also discuss how to interpret the results in cases of unknown identity and disputed parentage.-- Covers all steps from extraction of human DNA through to analysis and interpretation-- Takes advantage of new methodologies such as capillary electrophoresis-- Clear step-by-step instructions ensure unfailing reproducibility.


Plant Virology Protocols

Plant Virology Protocols

Author: Gary D. Foster

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 557

ISBN-13: 1592595669

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The aim of Plant Virology Protocols is to provide a source of infor- tion to guide the reader through the wide range of methods involved in gen- ating transgenic plants that are resistant to plant viruses. To this end, we have commissioned a wide-ranging list of chapters that will cover the methods required for: plant virus isolation; RNA extraction; cloning coat p- tein genes; introduction of the coat protein gene into the plant genome; and testing transgenic plants for resistance. The book then moves on to treatments of the mechanisms of resistance, the problems encountered with field testing, and key ethical issues surrounding transgenic technology. Although Plant Virology Protocols deals with the cloning and expression of the coat protein gene, the techniques described can be equally applied to other viral genes and nucleotide sequences, many of which have also been shown to afford protection when introduced into plants. The coat protein has, however, been the most widely applied, and as such has been selected to illustrate the techniques involved. Plant Virology Protocols has been divided into six major sections, c- taining 55 chapters in total.


Immunochemical Protocols

Immunochemical Protocols

Author: John Pound

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 501

ISBN-13: 1592592570

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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.