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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


Protein Structure Analysis

Protein Structure Analysis
Author: Roza Maria Kamp
Publisher: Springer Science & Business Media
Total Pages: 340
Release: 1997
Genre: Science
ISBN: 9783540615002

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"Protein Structure Analysis - Preparation and Characterization" is a compilation of practical approaches to the structural analysis of proteins and peptides. Here, about 20 authors describe and comment on techniques for sensitive protein purification and analysis. These methods are used worldwide in biochemical and biotechnical research currently being carried out in pharmaceu tical and biomedical laboratories or protein sequencing facilities. The chapters have been written by scientists with extensive ex perience in these fields, and the practical parts are well documen ted so that the reader should be able to easily reproduce the described techniques. The methods compiled in this book were demonstrated in student courses and in the EMBO Practical Course on "Microsequence Analysis of Proteins" held in Berlin September 10-15, 1995. The topics also derived from a FEBS Workshop, held in Halkidiki, Thessaloniki, Greece, in April, 1995. Most of the authors participated in these courses as lecturers and tutors and made these courses extremely lively and successful. Since polypeptides greatly vary depending on their specific structure and function, strategies for their structural analysis must for the most part be adapted to each individual protein. Therefore, advantages and limitations of the experimen tal approaches are discussed here critically, so that the reader becomes familiar with problems that might be encountered.


A Practical Guide to Membrane Protein Purification

A Practical Guide to Membrane Protein Purification
Author: Gebhard von Jagow
Publisher: Elsevier
Total Pages: 185
Release: 2013-10-22
Genre: Science
ISBN: 0080571727

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A Practical Guide to Membrane Protein Purification is written especially for researchers who have some familarity with separation of water-soluble proteins, but who may not be aware of the pitfalls they face with membrane proteins. This guide presents techniques in a concise form, emphasizing the aspects unique to membrane proteins. The book explains the principles of the methods, permitting researchers and students new to this area to adapt these techniques to their particular needs. The second volume in the series, this book is an essential manual for investigations of structure and function of native membrane proteins, as well as for purification of these proteins for immunization and protein sequencing. Separation, Detection, and Characterization of Biological Macromolecules is a new series of laboratory guides. Each volume focuses on a topic of central interest to scientists and students in biomedical and biological research. Introductory chapters are followed by clear, step-by-step protocols that present principles and practice. These concise manuals are designed for optimal understanding of methods as well as for practical benchtop use. Provides general guidelines and strategies for isolation of membrane proteins Describes detailed practical procedures that have been the widest applications, and lowest specialized equipment needs Gives special emphasis to new native and denaturing electrophoresis techniques Explains modifications of techniques used for water-soluble proteins


Protein Structure Analysis

Protein Structure Analysis
Author: Roza Maria Kamp
Publisher: Springer
Total Pages: 316
Release: 2012-11-07
Genre: Science
ISBN: 9783642477652

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"Protein Structure Analysis - Preparation and Characterization" is a compilation of practical approaches to the structural analysis of proteins and peptides. Here, about 20 authors describe and comment on techniques for sensitive protein purification and analysis. These methods are used worldwide in biochemical and biotechnical research currently being carried out in pharmaceu tical and biomedical laboratories or protein sequencing facilities. The chapters have been written by scientists with extensive ex perience in these fields, and the practical parts are well documen ted so that the reader should be able to easily reproduce the described techniques. The methods compiled in this book were demonstrated in student courses and in the EMBO Practical Course on "Microsequence Analysis of Proteins" held in Berlin September 10-15, 1995. The topics also derived from a FEBS Workshop, held in Halkidiki, Thessaloniki, Greece, in April, 1995. Most of the authors participated in these courses as lecturers and tutors and made these courses extremely lively and successful. Since polypeptides greatly vary depending on their specific structure and function, strategies for their structural analysis must for the most part be adapted to each individual protein. Therefore, advantages and limitations of the experimen tal approaches are discussed here critically, so that the reader becomes familiar with problems that might be encountered.


Gel Electrophoresis of Proteins

Gel Electrophoresis of Proteins
Author: B. D. Hames
Publisher: OUP Oxford
Total Pages: 378
Release: 1998-10-01
Genre: Science
ISBN: 0191565636

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This new edition of Gel Electrophoresis of Proteins is a completely new text, with eight of the ten chapters written by new authors. It presents the best methods, hints and tips for core procedures such as one- dimensional polyacrylamide gel electrophoresis, isoelectric focusing, two-dimensional gel electrophoresis, preparative gel electrophoresis, and peptide mapping, complete with the latest refinements and updates of the procedures. In addition, it describes major new techniques which have come to the fore since the previous edition. Thus there are chapters on capillary gel electrophoresis, sequence analysis of gel-resolved proteins, fluorophore-labelled saccharide electrophoresis, and analysis of protein:protein interactions by gel electrophoresis. One thing has not changed. The emphasis is still on describing the best methods, in step-by-step detail, with copious advice to ensure that each method works first time in the reader's hands. The first two editions of Gel Electrophoresis of Proteins: A Practical Approach each gained a strong reputation as easy-to-follow laboratory manuals written by experienced researchers for researchers. The methods were presented in a clear accessible format and had been fully tested to ensure success in the lab. This new edition will strengthen the reputation of the book still further. It is a 'must have' for all those who currently use gel electrophoresis or who plan to do so.


The Protein Protocols Handbook

The Protein Protocols Handbook
Author: John M. Walker
Publisher: Springer Science & Business Media
Total Pages: 759
Release: 2008-02-12
Genre: Science
ISBN: 1603272593

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In The Protein Protocols Handbook, I have attempted to provide a cross-section of analytical techniques commonly used for proteins and peptides, thus providing a benehtop manual and guide both for those who are new to the protein chemistry laboratory and for those more established workers who wish to use a technique for the first time. We each, of course, have our own favorite, commonly used gel system, g- staining method, blotting method, and so on; I'm sure you will find yours here. H- ever, I have also described a variety of altematives for many of these techniques; though they may not be superior to the methods you commonly use, they may nev- theless be more appropriate in a particular situation. Only by knowing the range of techniques that are available to you, and the strengths and limitations of these te- niques, will you be able to choose the method that best suits your purpose.


Methods of Protein Microcharacterization

Methods of Protein Microcharacterization
Author: John E. Shively
Publisher: Springer Science & Business Media
Total Pages: 458
Release: 2008-02-22
Genre: Science
ISBN: 1592594360

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Milestones in the techniques and methodology of polypeptide structure determination include the determination of the sequence of insulin by Sanger in 1951 (I) and the introduction of the repeti tive degradation of proteins with phenylisothiocyanate by Edman in 1959 (2). The automation of Edman chemistry (3) played a major role in the determination of polypeptide structures. Important modifications of Edman chemistry include the solid-phase approach by Laursen in 1971 (4) and the use of modified Edman reagents such as 4-N, N-dimethylaminoazobenzene-4'-isothiocy- ate (DABITC) for manual sequencing by Chang et al. (5) in 1976. A second major breakthrough in the analysis of polypeptides was automated amino acid analysis described by Spackman et al. in 1958 (6). However, during the period from 1975 to 1980, it became increasingly clear that the amount of material required for struc tural analysis was more than could be easily isolated for the vast majority of proteins. The field was criticized for its lack of sensitive techniques for the analysis of growth factors, immune modulators, membrane receptors, and peptide hormones. In addition, very little had been done to modernize and improve the original instruments introduced in the mid-1960s. The first indications of improved instrumentation for Edman chemistry came from Wittmann-Liebold's laboratory (7), followed by the introduction of a "micro" sequencer by Hunkapiller and Hood in 1978 (8). The movement toward improved instrumentation culminated in the "gas"--Phase sequencer of Hewick et al. (9) in 1981


Protein Purification Applications

Protein Purification Applications
Author: Simon Roe
Publisher: OUP Oxford
Total Pages: 182
Release: 2001-01-25
Genre: Science
ISBN: 0191565733

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Proteins are an integral part of molecular and cellular structure and function and are probably the most purified type of biological molecule. In order to elucidate the structure and function of any protein it is first necessary to purify it. Protein purification techniques have evolved over the past ten years with improvements in equipment control, automation, and separation materials, and the introduction of new techniques such as affinity membranes and expanded beds. These developments have reduced the workload involved in protein purification, but there is still a need to consider how unit operations linked together to form a purification strategy, which can be scaled up if necessary. The two Practical Approach books on protein purification have therefore been thoroughly updated and rewritten where necessary. The core of both books is the provision of detailed practical guidelines aimed particularly at laboratory scale purification. Information on scale-up considerations is given where appropriate. The books are not comprehensive but do cover the major laboratory techniques and common sources of protein. Protein Purification Techniques focuses on unit operations and analytical techniques. It starts with an overview of purification strategy and then covers initial extraction and clarification techniques. The rest of the book concentrates on different purification methods with the emphasis being on chromatography. The final chapter considers general scale-up considerations. Protein Purification Applications describes purification strategies from common sources: mammalian cell culture, microbial cell culture, milk, animal tissue, and plant tissue. It also includes chapters on purification of inclusion bodies, fusion proteins, and purification for crystallography. A purification strategy that can produce a highly pure single protein from a crude mixture of proteins, carbohydrates, lipids, and cell debris to is a work of art to be admired. These books (available individually or as a set)are designed to give the laboratory worker the information needed to undertake the challenge of designing such a strategy.


Guide to Protein Purification

Guide to Protein Purification
Author: Richard R Burgess
Publisher: Academic Press
Total Pages: 915
Release: 2009-11-03
Genre: Science
ISBN: 0080923178

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Guide to Protein Purification, Second Edition provides a complete update to existing methods in the field, reflecting the enormous advances made in the last two decades. In particular, proteomics, mass spectrometry, and DNA technology have revolutionized the field since the first edition’s publication but through all of the advancements, the purification of proteins is still an indispensable first step in understanding their function. This volume examines the most reliable, robust methods for researchers in biochemistry, molecular and cell biology, genetics, pharmacology and biotechnology and sets a standard for best practices in the field. It relates how these traditional and new cutting-edge methods connect to the explosive advancements in the field. This "Guide to" gives imminently practical advice to avoid costly mistakes in choosing a method and brings in perspective from the premier researchers while presents a comprehensive overview of the field today. Gathers top global authors from industry, medicine, and research fields across a wide variety of disciplines, including biochemistry, genetics, oncology, pharmacology, dermatology and immunology Assembles chapters on both common and less common relevant techniques Provides robust methods as well as an analysis of the advancements in the field that, for an individual investigator, can be a demanding and time-consuming process


Methods for Protein Analysis

Methods for Protein Analysis
Author: Robert A. Copeland
Publisher: Springer Science & Business Media
Total Pages: 238
Release: 2013-11-11
Genre: Science
ISBN: 1475715056

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As protein science continues to become an increasingly important aspect of academic and commercial sciences and technology, the need has arisen for a ready source of laboratory protocols for the analysis and evaluation of these biological polymers. Methods for Protein Analysis presents the methods most relevant to the generalist bench scientist working with proteins. A concise yet thorough summary, it covers laboratory methods that can be reasonably performed in a standard protein laboratory, without specialized equipment or expertise. Taking a how to approach, this book examines the techniques used to answer common protein analytical questions and describes methods useful in daily laboratory work. Methods for Protein Analysis is the ideal reference for protein laboratories in academic, government and industrial settings. It is an essential benchtop manual for first-year graduate students beginning their laboratory experience as well as for chemists, biochemists, and molecular biologists in the pharmaceutical, biotechnological, food and specialty chemical industries, and for analysts concerned with the purity and structural integrity of protein. Featuring illustrations and a convenient spiral binding, this guide offers a glossary of common abbreviations and a list of suppliers for protein science.