Methods In Molecular Biology Protein Purification Protocols
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| Author | : Shawn Doonan |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 406 |
| Release | : 2008-02-02 |
| Genre | : Science |
| ISBN | : 1592595456 |
Hans Neurath has written that this is the second golden era of enzymology {Protein Science [1994], vol. 3, pp. 1734—1739); he could with justice have been more general and referred to the second golden age of protein chemistry. The last two decades have seen enormous advances in our understanding of the structures and functions of pro teins arising on the one hand from improvements and developments in analytical techniques {see the companion volume, Basic Protein and Peptide Protocols, in this series) and on the other hand from the tech nologies of molecular genetics. Far from turning the focus away from protein science, the ability to isolate, analyze, and express genes has increased interest in proteins as gene products. Hence, many laborato ries are now getting involved in protein isolation for the first time, either as an essential adjunct to their work in molecular genetics or because of a curiosity to know more about the products of the genes that they have been studying. Protein Purification Protocols is aimed mainly at these newcom ers to protein purification, but it is hoped that it will also be of value to established practitioners who may find here techniques that they have not tried, but which might well be most applicable in their work. With the exception mainly of the first and last chapters, the format of the contributions to the present book conform to the established format of the Methods in Molecular Biology series.
| Author | : Paul Cutler |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 474 |
| Release | : 2008-02-02 |
| Genre | : Science |
| ISBN | : 159259655X |
The first edition of Protein Purification Protocols (1996), edited by Professor Shawn Doonan, rapidly became very successful. Professor Doonan achieved his aims of p- ducing a list of protocols that were invaluable to newcomers in protein purification and of significant benefit to established practitioners. Each chapter was written by an ex- rienced expert in the field. In the intervening time, a number of advances have w- ranted a second edition. However, in attempting to encompass the recent developments in several areas, the intention has been to expand on the original format, retaining the concepts that made the initial edition so successful. This is reflected in the structure of this second edition. I am indebted to Professor Doonan for his involvement in this new edition and the continuity that this brings. Each chapter that appeared in the original volume has been reviewed and updated to reflect advances and bring the topic into the 21st century. In many cases, this reflects new applications or new matrices available from vendors. Many of these have increased the performance and/or scope of the given method. Several new chapters have been introduced, including chapters on all the currently used protein fractionation and ch- matographic techniques. They introduce the theory and background for each method, providing lists of the equipment and reagents required for their successful execution, as well as a detailed description of how each is performed.
| Author | : Barry S. Selinsky |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 330 |
| Release | : 2008-02-03 |
| Genre | : Science |
| ISBN | : 159259400X |
Knowledge of the three-dimensional structure of a protein is absolutely required for the complete understanding of its function. The spatial orientation of amino acids in the active site of an enzyme demonstrates how substrate specificity is defined, and assists the medicinal chemist in the design of s- cific, tight-binding inhibitors. The shape and contour of a protein surface hints at its interaction with other proteins and with its environment. Structural ana- sis of multiprotein complexes helps to define the role and interaction of each individual component, and can predict the consequences of protein mutation or conditions that promote dissociation and rearrangement of the complex. Determining the three-dimensional structure of a protein requires milligram quantities of pure material. Such quantities are required to refine crystallization conditions for X-ray analysis, or to overcome the sensitivity limitations of NMR spectroscopy. Historically, structural determination of proteins was limited to those expressed naturally in large amounts, or derived from a tissue or cell source inexpensive enough to warrant the use of large quantities of cells. H- ever, with the advent of the techniques of modern gene expression, many p- teins that are constitutively expressed in minute amounts can become accessible to large-scale purification and structural analysis.
| Author | : John M. Walker |
| Publisher | : |
| Total Pages | : |
| Release | : 1984 |
| Genre | : Molecular biology -- v.236. Plant functional genomics -- v.237. G protein signaling -- v.238. Biopolymer methods in tissue engineering -- v.239. Cell migration in inflammation and immunity -- v.240. Mammalian artificial chromosomes -- v.241. Cell cycle checkpoint control protocols -- v.242. Atomic force microscopy -- v.243. Chiral separations: Methods and protocols -- v.244. Protein purification protocols -- v.245-6. Gene delivery to mammalian cells -- v.247. Drosophila cytogenetics protocols -- v.248. Antibody engineering -- v.249. Cytokine protocols -- v.251. HPLC of peptides and proteins: Methods and protocols -- v.265. RNA interference, editing, and modification -- v.274. Photosynthesis research protocols -- v.318. Plant cell culture protocols -- v.323. Arabidopsis protocols |
| ISBN | : |
| Author | : Dermot Walls |
| Publisher | : Humana Press |
| Total Pages | : 527 |
| Release | : 2011-04-06 |
| Genre | : Science |
| ISBN | : 9781607619147 |
A prerequisite for elucidating the structure and function of any protein is the prior purification of that protein. This necessity has led to the development of many purification schemes and chromatographic methods for the isolation of native proteins from complex sources. In Protein Chromatography: Methods and Protocols, leading researchers present clear protocol-style chapters that are suitable for newcomers and experts alike. The book opens with vital topics in protein biochemistry, addressing such areas as protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the contrasting challenges that microfluidics and scale-up production pose to the investigator, and then it segues into key methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, amongst other means, as well as many variations on classic techniques such as ion-exchange and immunoaffinity chromatography. Written in the highly successful Methods in Molecular BiologyTM series format, protocols chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Protein Chromatography: Methods and Protocols will greatly aid scientists in establishing these essential techniques in their own laboratories and furthering our understanding of the many imperative functions of proteins.
| Author | : Sinéad T Loughran |
| Publisher | : Springer Nature |
| Total Pages | : 495 |
| Release | : 2023-10-01 |
| Genre | : Science |
| ISBN | : 1071633627 |
This third edition expands on the previous editions with updated and new chapters on protein chromatography. Chapters detail protein stability and storage, avoiding proteolysis, protein quantitation methods, generation and purification of recombinant proteins, recombinant antibody production, and the tagging of proteins. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Protein Chromatography: Methods and Protocols, Third Edition aims to provide commonly used methods and new approaches to help both new researchers and experts expand their knowledge.
| Author | : Paul Cutler |
| Publisher | : |
| Total Pages | : 498 |
| Release | : 2004 |
| Genre | : Proteins |
| ISBN | : 9781489939432 |
In this new edition of the very successful Protein Purification Protocols (1996), Paul Cutler completely updates the existing protocols to reflect recent advances and adds an enormous new array of proteomic techniques for protein isolation and analysis. These cutting-edge techniques include not only two-dimensional gel electrophoresis for analysis and characterization, but also analytical chromatography for multidimensional separations of proteins and peptides, and mass spectrometry for isolating proteins. With the many recent advances in technology, simple spectrometric detection is no longer the only option for separating proteins, and the authors treat in full detail all the newer methods for these separations. Comprehensive and highly practical, Protein Purification Protocols, Second Edition, brings together all the key methodologies that both novice and experienced investigators need to carry out successful experimental work on proteins and their functions today.
| Author | : Isabelle Mus-Veteau |
| Publisher | : Springer Nature |
| Total Pages | : 472 |
| Release | : 2022-06-30 |
| Genre | : Science |
| ISBN | : 1071623680 |
This detailed volume explores protocols for the production of membrane proteins in a panel of heterologous organisms for structural studies. Beginning with techniques using E. coli as a host for the overproduction and purification of membrane proteins, the book continues with chapters covering mammalian membrane protein production in yeast, insect cells, mammalian cells, as well as using virus like particles and acellular systems. Additionally, new detergents and alternatives to detergents allowing membrane protein purification for structural analyses are described. The book closes with a chapter exploring the use of microscale thermophoresis (MST) to evaluate the binding activity of heterologously expressed proteins directly in crude membrane extracts. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Heterologous Expression of Membrane Proteins: Methods and Protocols, Third Edition serves as an ideal guide for scientists aiming to produce and purify functional recombinant membrane proteins for structural studies.
| Author | : Elena Garcia Fruitós |
| Publisher | : Humana |
| Total Pages | : 0 |
| Release | : 2023-02-12 |
| Genre | : Science |
| ISBN | : 9781071618615 |
This volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Chapters describe recombinant protein production in different expression systems, prokaryotic and eukaryotic expression systems, purification protocols, characterization of insoluble proteins and a general overview of interesting applications of insoluble proteins. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Insoluble Proteins: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.
| Author | : Renaud Vincentelli |
| Publisher | : Humana |
| Total Pages | : 537 |
| Release | : 2019-07-05 |
| Genre | : Science |
| ISBN | : 9781493996230 |
This book compiles key protocols instrumental to the study of high-throughput protein production and purification which have been refined and simplified over the years and are now ready to be transferred to any laboratory. Beginning with a section covering general procedures for high-throughput protein production, the volume continues with high-throughput protocols adapted to the production of specific protein families, as well as an extensive section on protocols combining high-throughput protein production and their micro-characterization. Written for the highly successful Methods in Molecular Biology series, chapters in this book include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, High-Throughput Protein Production and Purification: Methods and Protocols serves biochemists ranging from engineers, PhD students and post-doctoral fellows, to the heads of protein expression facilities and researchers, in pursuing this vital area of study.
