Gel Electrophoresis of Proteins: A Practical Approach (Practical Approach Series) PDF
Preview Gel Electrophoresis of Proteins: A Practical Approach (Practical Approach Series)
Gel Electrophoresis of Proteins The Practical Approach Series SERIES EDITOR B. D. HAMES School of Biochemistry and Molecular Biology University of Leeds, Leeds LS2 9JT, UK See also the Practical Approach web site at http://www.oup.co.uk/PAS * indicates new and forthcoming titles Affinity Chromatography * Cell Separation if Affinity Separations Cellular Calcium Anaerobic Microbiology Cellular Interactions in Development Animal Cell Culture (2nd edition) Cellular Neurobiology Animal Virus Pathogenesis * Chromatin Antibodies I and II Clinical Immunology Antibody Engineering if Complement Antisense Technology Crystallization of Nucleic Acids and Proteins if Applied Microbial Physiology Cytokines (2nd edition) Basic Cell Culture The Cytoskeleton Behavioural Neuroscience Diagnostic Molecular Bioenergetics Pathology I and II Biological Data Analysis DNA and Protein Sequence Biomaterial Immobilization Analysis Biomechanics—Materials DNA Cloning 1: Core Biomechanics—Structures and Techniques (2nd edition) Systems DNA Cloning 2: Expression Biosensors Systems (2nd edition) Carbohydrate Analysis (2nd DNA Cloning 3: Complex edition) Genomes (2nd edition) Cell-Cell Interactions DNA Cloning 4: Mammalian The Cell Cycle Systems (2nd edition) Cell Growth and Apoptosis Drosophila (2nd edition) Electron Microscopy in Immunochemistry 2 Biology Immunocytochemistry Electron Microscopy in * In Situ Hybridization (2nd Molecular Biology edition) Electrophysiology lodinated Density Gradient Enzyme Assays Media Epithelial Cell Culture Ion Channels Essential Developmental if Light Microscopy (2nd edition) Biology Lipid Modification of Proteins Essential Molecular Biology I Lipoprotein Analysis and II Liposomes Experimental Neuroanatomy Mammalian Cell Extracellular Matrix Biotechnology Flow Cytometry (2nd edition) Medical Parasitology Free Radicals Medical Virology Gas Chromatography * MHC Volumes 1 and 2 Gel Electrophoresis of Nucleic if Molecular Genetic Analysis of Acids (2nd edition) Populations (2nd edition) Gel Electrophoresis of Molecular Genetics of Yeast Proteins (3rd edition) Molecular Imaging in Gene Probes 1 and 2 Neuroscience Gene Targeting Molecular Neurobiology Gene Transcription Molecular Plant Pathology I and II * Genome Mapping Molecular Virology Glycobiology Monitoring Neuronal Activity * Growth Factors and Receptors Mutagenicity Testing Haemopoiesis if Mutation Detection Histocompatibility Testing Neural Cell Culture HIV Volumes land 2 Neural Transplantation HPLC of Macromolecules (2nd Neurochemistry (2nd edition) edition) Neuronal Cell Lines Human Cytogenetics I and II NMR of Biological (2nd edition) Macromolecules Human Genetic Disease Non-isotopic Methods in Analysis Molecular Biology Immunochemistry 1 Nucleic Acid Hybridization Oligonucleotides and Analogues Protein Function (2nd edition) Oligonucleotide Synthesis Protein Phosphorylation PCR1 Protein Purification Applications PCR2 Protein Purification Methods Hybridization Protein Sequencing Peptide Antigens Protein Structure (2nd edition) Photosynthesis: Energy Protein Structure Prediction Transduction Protein Targeting Plant Cell Biology Proteolytic Enzymes Plant Cell Culture (2nd edition) Pulsed Field Gel Plant Molecular Biology Electrophoresis Plasmids (2nd edition) RNA Processing I and II Platelets * RNA-Protein Interactions Postimplantation Mammalian * Signalling by Inositides Embryos Subcellular Fractionation Preparative Centrifugation Signal Transduction Protein Blotting Transcription Factors Protein Engineering Tumour Immunobiology Gel Electrophoresis of Proteins A Practical Approach THIRD EDITION Edited by B. D. HAMES School of Biochemistry and Molecular Biology University of Leeds, Leeds LS2 9JT, UK Oxford New York Tokyo OXFORD UNIVERSITY PRESS 1998 Oxford University Press, Great Clarendon Street, Oxford OX2 6DP Oxford New York Athens Auckland Bangkok Bogota Bombay Buenos Aires Calcutta Cape Town Dares Salaam Delhi Florence Hong Kong Istanbul Karachi Kuala Lumpur Madrid Melbourne Mexico City Mumbai Nairobi Paris Sao Paolo Singapore Taipei Tokyo Toronto Warsaw and associated companies in Berlin Ibadan Oxford is a trade mark of Oxford University Press Published in the United States by Oxford University Press Inc., New York © Oxford University Press, 1998 All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, without the prior permission in writing of Oxford University Press. Within the UK, exceptions are allowed in respect of any fair dealing for the purpose of research or private study, or criticism or review, as permitted under the Copyright, Designs and Patents Act, 1988, or in the case of reprographic reproduction in accordance with the terms of licences issued by the Copyright Licensing Agency. Enquiries concerning reproduction outside those terms and in other countries should be sent to the Rights Department, Oxford University Press, at the address above. This book is sold subject to the condition that it shall not, by way of trade or otherwise, be lent, re-sold, hired out, or otherwise circulated without the publisher's prior consent in any form of binding or cover other than that in which it is published and without a similar condition including this condition being imposed on the subsequent purchaser. Users of books in the Practical Approach Series are advised that prudent laboratory safety procedures snould be followed at all times. Oxford University Press makes no representation, express or implied, in respect of the accuracy of the material set forth in books in this series and cannot accept any legal responsibility or liability for any errors or omissions that may be made. A catalogue record for this book is available from the British Library Library of Congress Cataloging in Publication Data Gel electrophoresis of proteins: a practical approach/edited by B.D. Homes.—3rd ed. (Practical approach series; 197) Includes bibliographical references and index. 1. Proteins—Analysis. 2. Gel electrophoresis. I. Homes, B.D. II. Series. QP551.G334 1998 572'.6—dc21 98-18884 CIP ISBN 0 19 963641 9 (Hbk) 0 19 963640 0 (Pbk) Typeset by Footnote Graphics, Warminster, Wilts Printed in Great Britain by Information Press Ltd., Eynsham, Oxon Preface When the first edition of this book was published in 1981,1 had no idea that it would lead to a major series of laboratory manuals, the Practical Approach series, currently over 190 volumes in size and still growing. The popularity of the series has led to second editions of many books. The second edition of Gel Electrophoresls of Proteins: A Practical Approach was duly published in 1990. Since that time, the field has continued to develop apace, so much so that some current major techniques, such as capillary electrophoresis, were not even mentioned in the second edition. Clearly, it is time for a third edition! The goal in devising and editing the third edition of Gel Electrophoresis of Proteins: A Practical Approach has been to reflect the laboratory usage of the wide variety of methods that fall under this general subject heading. I make no apology for revisiting the basic techniques of one-dimensional polyacrylamide gel electrophoresis, including SDS-PAGE, since these methods have contin- ued to be widely used on a regular basis in enormous numbers of laboratories around the world, but even here there have been some very useful develop- ments in technology. These are fully described in the first chapter which has been written by new authors to bring fresh insights to the subject. Other tech- niques have also undergone major developments in the intervening years, as will be clear to readers of the chapters on isoelectric focusing, two-dimensional gel electrophoresis, preparative gel electrophoresis, and polypeptide detection systems. However, my aim was also to cover important emerging techniques. Thus there are completely new chapters on capillary gel electrophoresis, sequence analysis of gel-resolved proteins, fluorophore-labelled saccharide electrophoresis, and analysis of protein:protein interactions by gel electro- phoresis. In total, only four of the chapter subjects covered in the previous edition are retained in the ten chapters of this new edition. Furthermore, eight of the ten chapters in this book are written by new authors. Thus the third edi- tion is in every sense a new text, and very definitely a major update in the field. I am greatly indebted to the authors of each chapter who accepted with good grace the very large number of editing changes I requested in that elusive quest for perfection. Given that the authors are active researchers with intimate knowledge of the methods (and pitfalls!) they describe, I am certain that the book will help colleagues to apply the new methods successfully in their own work. However, I have no illusions that this is the end of the story. The versatility and power of gel electrophoretic techniques in biological research ensure that still further developments and applications will continue to arise in future. Leeds January 1998 B. David Hames This page intentionally left blank Contents List of contributors xvii Abbreviations xix 1. One-dimensional polyacrylamide gel electrophoresis 1 Qinwei Shi and George Jackowski 1. Introduction 1 2. The polyacrylamide gel matrix 2 Chemical structure and mechanism of polymerization 2 Factors affecting polymerization 3 Resolving range of polyacrylamide gels 7 3. Gel apparatus 8 Electrophoresis apparatus 8 Power pack 10 Apparatus for gradient gels 10 Photopolymerization equipment 10 4. Choice of a gel system 11 Non-dissociating systems 11 Dissociating systems 11 Transverse gradient gels 13 5. SDS-polyacrylamide gel electrophoresis 13 Introduction 13 Reagent preparation 14 Gel preparation 16 Sample preparation 21 Sample loading and electrophoresis 27 Molecular mass estimation 29 Troubleshooting 33 6. Non-denaturing polyacrylamide gel electrophoresis 35 Introduction 35 Choice of buffers and polymerization catalyst 37 Preparation of non-denaturing slab gels 38 Sample preparation and electrophoresis of native proteins 39 7. Variations of standard polyacrylamide gel electrophoresis 40 Blue native polyacrylamide gel electrophoresis 40 Acid-urea polyacrylamide gel electrophoresis 42 Transverse gradient gel electrophoresis 42
Description:The list of books you might like
