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Protein Sequencing Protocols

Protein Sequencing Protocols
Author: Bryan John Smith
Publisher: Springer Science & Business Media
Total Pages: 489
Release: 2008-02-02
Genre: Science
ISBN: 1592593429

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Determination of the protein sequence is as important today as it was a half century ago, even though the techniques and purposes have changed over time. Mass spectrometry has continued its recent rapid development to find notable application in the characterization of small amounts of protein, for example, in the field of proteomics. The “traditional” chemical N-terminal sequencing is still of great value in quality assurance of the increasing number of biopharmaceuticals that are to be found in the clinic, checking processing events of recombinant proteins, and so on. It is joined in the armory of me- ods of protein analysis by such techniques as C-terminal sequencing and amino acid analysis. These methods are continually developing. The first edition of Protein Sequencing Protocols was a “snapshot” of methods in use in protein biochemistry laboratories at the time, and this, the second edition, is likewise. Methods have evolved in the intervening period, and the content of this book has similarly changed, the content of some chapters having been superceded and replaced by other approaches. Thus, in this edition, there is inclusion of approaches to validation of methods for quality assurance work, reflecting the current importance of biopharmaceuticals, and also a guide to further analysis of protein sequence information, acknowledging the importance of bioinformatics.


Protein Sequencing and Identification Using Tandem Mass Spectrometry

Protein Sequencing and Identification Using Tandem Mass Spectrometry
Author: Michael Kinter
Publisher: John Wiley & Sons
Total Pages: 321
Release: 2005-04-12
Genre: Science
ISBN: 0471231886

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How to design, execute, and interpret experiments for protein sequencing using mass spectrometry The rapid expansion of searchable protein and DNA databases in recent years has triggered an explosive growth in the application of mass spectrometry to protein sequencing. This timely and authoritative book provides professionals and scientists in biotechnology research with complete coverage of procedures for analyzing protein sequences by mass spectrometry, including step-by-step guidelines for sample preparation, analysis, and data interpretation. Michael Kinter and Nicholas Sherman present their own high-quality, laboratory-tested protocols for the analysis of a wide variety of samples, demonstrating how to carry out specific experiments and obtain fast, reliable results with a 99% success rate. Readers will get sufficient experimental detail to apply in their own laboratories, learn about the proper selection and operation of instruments, and gain essential insight into the fundamental principles of mass spectrometry and protein sequencing. Coverage includes: * Peptide fragmentation and interpretation of product ion spectra * Basic polyacrylamide gel electrophoresis * Preparation of protein digests for sequencing experiments * Mass spectrometric analysis using capillary liquid chromatography * Techniques for protein identification by database searches * Characterization of modified peptides using tandem mass spectrometry And much more


A Practical Guide to Protein and Peptide Purification for Microsequencing

A Practical Guide to Protein and Peptide Purification for Microsequencing
Author: Paul T. Matsudaira
Publisher: Elsevier
Total Pages: 205
Release: 2012-12-02
Genre: Science
ISBN: 0080924611

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Why a Second Edition? The Second Edition provides practical answers to the general question, "How can I obtain useful sequence information from my protein or peptide?" rather than the more specific question asked in the first edition, "How can I obtain the N-terminal sequence?" Important new methods include ways of dealing with blocked N termini, computer analysis of protein sequences, and the recent revolution in mass spectrometry. Mass spectrophotometric characterization of proteins and peptides N-terminal sequencing of proteins with blocked N termini Internal amino acid sequence analysis after protease digestion in-gel and on-blot Improved microscale peptide purification methods Computer analysis of protein sequences New protocols tested and refined through everyday use in authors' laboratories Updated reference chapter covering all aspects of protein microsequencing


2-D Proteome Analysis Protocols

2-D Proteome Analysis Protocols
Author: Andrew J. Link
Publisher: Springer Science & Business Media
Total Pages: 585
Release: 2008-02-02
Genre: Science
ISBN: 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.


Protein Engineering Protocols

Protein Engineering Protocols
Author: Kristian Müller
Publisher: Springer Science & Business Media
Total Pages: 318
Release: 2007-10-26
Genre: Science
ISBN: 1597451878

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Protein engineering is a fascinating mixture of molecular biology, protein structure analysis, computation, and biochemistry, with the goal of developing useful or valuable proteins. Protein Engineering Protocols will consider the two general, but not mutually exclusive, strategies for protein engineering. The first is known as rational design, in which the scientist uses detailed knowledge of the structure and function of the protein to make desired changes. The s- ond strategy is known as directed evolution. In this case, random mutagenesis is applied to a protein, and selection or screening is used to pick out variants that have the desired qualities. By several rounds of mutation and selection, this method mimics natural evolution. An additional technique known as DNA shuffling mixes and matches pieces of successful variants to produce better results. This process mimics recombination that occurs naturally during sexual reproduction. The first section of Protein Engineering Protocols describes rational p- tein design strategies, including computational methods, the use of non-natural amino acids to expand the biological alphabet, as well as impressive examples for the generation of proteins with novel characteristics. Although procedures for the introduction of mutations have become routine, predicting and und- standing the effects of these mutations can be very challenging and requires profound knowledge of the system as well as protein structures in general.


Short Protocols in Protein Science

Short Protocols in Protein Science
Author: John E. Coligan
Publisher: Wiley-Blackwell
Total Pages: 832
Release: 2003-10-24
Genre: Science
ISBN:

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Short Protocols in Protein Science provides condensed descriptions of more than 500 protocols compiled from Current Protocols in Protein Science. Drawing from both the original "core" manual as well as the quarterly update service, this compendium includes all step-by-step descriptions of the principal methods covered in Current Protocols in Protein Science.


Protein Sequencing

Protein Sequencing
Author: Michael J. Geisow
Publisher: Irl Press
Total Pages: 199
Release: 1989
Genre: Amino acid sequence
ISBN: 9780199630134

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Here is a convenient laboratory manual on protein sequence analysis, an area currently at the forefront of modern molecular biology. The volume describes how to isolate proteins or peptides, and provides information on subsequent sequence analysis and interpretation. Written for both professional and inexperienced research workers, and for those with and without access to highly technology instrumentation, readers will find this title to be a unique and highly useful source for the latest experimental methods and laboratory protocols. those wishing to isolate proteins or peptides, as well as providing a needed source of information on analysis and interpretations of structural significance. Written for both professional and inexperienced research workers, and for those with and without access to high technology instrumentation, readers will find this title to be a unique and highly informative source for the latest experimental methods and laboratory protocols.


The Protein Protocols Handbook

The Protein Protocols Handbook
Author: John M. Walker
Publisher: Springer Science & Business Media
Total Pages: 1422
Release: 2002
Genre: Medical
ISBN: 0896039404

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The authors are commonly the techniques" originators, and each has demonstrated a hands-on mastery of the methods described, always fine-tuning them here for optimal productivity.


Therapeutic Proteins

Therapeutic Proteins
Author: C. Mark Smales
Publisher: Springer Science & Business Media
Total Pages: 481
Release: 2008-02-04
Genre: Science
ISBN: 1592599222

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With the recent completion of the sequencing of the human genome, it is widely anticipated that the number of potential new protein drugs and targets will escalate at an even greater rate than that observed in recent years. However, identification of a potential target is only part of the process in developing these new next generation protein-based “drugs” that are increasingly being used to treat human disease. Once a potential protein drug has been identified, the next rate-limiting step on the road to development is the production of sufficient authentic material for testing, charact- ization, clinical trials, and so on. If a protein drug does actually make it through this lengthy and costly process, methodology that allows the production of the protein on a scale large enough to meet demand must be implemented. Furthermore, large-scale production must not compromise the authenticity of the final product. It is also nec- sary to have robust methods for the purification, characterization, viral inactivation and continued testing of the authenticity of the final protein product and to be able to formulate it in a manner that retains both its biological activity and lends itself to easy administration. Therapeutic Proteins: Methods and Protocols covers all aspects of protein drug production downstream of the discovery stage. This volume contains contributions from leaders in the field of therapeutic protein expression, purification, characterization, f- mulation, and viral inactivation.