Gel Electrophoresis Of Proteins

Gel Electrophoresis of Proteins focuses on the techniques, methodologies, reactions, and approaches involved in gel electrophoresis of proteins. The selection first covers steady-state gel electrophoresis systems and one-dimensional PAA-gel electrophoretic techniques to separate functional and denatured proteins. Discussions focus on affinity electrophoresis, structure and physico-chemical properties of polyacrylamide gels, moving boundary electrophoresis, isotachophoresis, fundamental steady-state electrophoresis systems, and fundamental properties of steady-state electrophoresis systems. The text then reviews conventional isoelectric focusing and immobilized pH gradients and high resolution two-dimensional polyacrylamide-gel electrophoresis. Topics include production of narrow pH gradients, extended pH gradients, polymerization kinetics, IEF in agarose matrices, titration curves, two-dimensional electrophoresis under non-denaturing conditions, and IEF in polyacrylamide matrices. The book tackles quantifying patterns from two-dimensional PAGE, protein staining and detection methods, and immunoelectrophoretic methods. Concerns include post-electrophoretic organic protein stains, silver and enzyme stains, detection of radioactive proteins, and programming languages. The selection is highly recommended for researchers wanting to conduct studies on gel electrophoresis of proteins.

Two-Dimensional Gel Electrophoresis of Proteins: Methods and Applications reviews current methods and clinical applications of two-dimensional gel electrophoresis of proteins, including the QUEST system, silver staining, and peptide mapping. Two-dimensional gel electrophoresis are applied to the study of diseases ranging from inborn errors of metabolism to human germ-line mutation rates, cancer, and mistranslation in animal and bacterial cells. This volume is organized into three sections encompassing 14 chapters and begins with an overview of the methodology of two-dimensional gel electrophoresis, followed by a discussion of computerized two-dimensional gel electrophoresis, silver staining, immunoblotting, and one- and two-dimensional peptide mapping. In most cases, a step-by-step guide to the techniques is given so that procedures may be easily repeated. A catalog of mouse fibroblast proteins is also given. The chapters that follow focus mainly on applications of two-dimensional gel electrophoresis in areas such as clinical and cancer research, human genetics, protein biosynthesis, and gene expression in plants. The final section presents current protein catalogs of Escherichia coli and human HeLa cells. This book is suitable for young researchers as well as for senior scientists working with a wide variety of problems in molecular and cell biology, basic biochemistry, genetics, and clinical research.

Proteins are the functional units of the cellular machinery and they provide significant information regarding the molecular basis of health and disease. Therefore, techniques to separate and isolate the various proteins are critical to studying and understanding their functional characteristics. One of the widely used techniques for this purpose is electrophoresis. In Protein Electrophoresis: Methods and Protocols, contributions from experts in the field have been collected in order to provide practical guidelines to this complex study. Each chapter outlines a specific electrophoretic variant in detail so that laboratory scientists may perform a technique new to their lab without difficulty. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Protein Electrophoresis: Methods and Protocols seeks to serve laboratory scientists with well-honed, detailed methodologies in an effort to further our knowledge of this essential field.

Immobilized pH gradients (IPG) represent the most advanced development of isoelectric focusing (IEF). Originally developed to overcome all the problems of IEF in soluble amphoteric buffers (CA) (such as pH gradient instability, complexation with CA chemicals, unreproducibility of pH gradients, protein precipitation at the pI), it turned out to be an entirely new technique, quite different in principle and operation from conventional IEF. The book is thus meant to bring the reader up to date with this fast developing field. The book is divided into six chapters containing information on: detailed treatment of all the chemistry of the Immobiline chemicals; theory of pH gradient generation (computer simulations, tables with all the possible pH recipes); all analytical aspects of IPGs, including staining techniques, blotting etc.; two-dimensional maps, with a detailed treatise of advantages and limitations; preparative aspects of IPGs, including comparison with other preparative electrophoretic techniques; some examples of applications, including genetic and forensic analysis, blood polymorphism etc.The book is extensive and up-to-date, while also extensively covering the theory. Clearly written, with easily accessible information, the volume is a worthy asset to the Lab. Techniques series.

The Protein Protocols Handbook, Second Edition aims to provide a cross-section of analytical techniques commonly used for proteins and peptides, thus providing a benchtop manual and guide 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. All chapters are written in the same format as that used in the Methods in Molecular BiologyTM series. Each chapter opens with a description of the basic theory behind the method being described. The Materials section lists all the chemicals, reagents, buffers, and other materials necessary for carrying out the protocol. Since the principal goal of the book is to provide experimentalists with a full account of the practical steps necessary for carrying out each protocol successfully, the Methods section contains detailed st- by-step descriptions of every protocol that should result in the successful execution of each method. The Notes section complements the Methods material by indicating how best to deal with any problem or difficulty that may arise when using a given technique, and how to go about making the widest variety of modifications or alterations to the protocol. Since the first edition of this book was published in 1996 there have, of course, been significant developments in the field of protein chemistry.