Protein Blotting and Detection Methods and Protocols

Over the past thirty years, the development of the Western blot has revolutionized the fields of biomedical research and medical diagnostics. In Protein Blotting and Detection: Methods and Protocols, expert researchers present numerous techniques based on

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IN

MOLECULAR BIOLOGY™

Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK

For other titles published in this series, go to www.springer.com/series/7651

METHODS

IN

MOLECULAR BIOLOGY™

Protein Blotting and Detection Methods and Protocols

Edited by

Biji T. Kurien and R. Hal Scofield Oklahoma Medical Research Foundation, Oklahoma City, OK, USA

Editors Biji T. Kurien Oklahoma Medical Research Foundation Oklahoma City OK, USA

R. Hal Scofield Oklahoma Medical Research Foundation Oklahoma City OK, USA

ISSN: 1064-3745 e-ISSN: 1940-6029 ISBN: 978-1-934115-73-2 e-ISBN: 978-1-59745-542-8 DOI: 10.1007/978-1-59745-542-8 Springer Dordrecht Heidelberg London New York Library of Congress Control Number: 2008940984 © Humana Press, a part of Springer Science+Business Media, LLC 2009 All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013 USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights. Cover illustration: Figure 15 in Chapter 26 Printed on acid-free paper springer.com

Preface Amazingly for a procedure so fundamental to present-day biomedical research as well as medical diagnostics, development of the western blot occurred within the memory of many individuals who continue to work in these fields. That is, conversely, there are many investigators and clinicians presently working who recall the days prior to the western blot. The personal stories of how and why the technique was developed are given in this volume by the people intimately involved with this development. These stories date from only roughly 30 years ago. Since then, a very large number of variations on the theme of gel separation of biologic products by different parameters such as size or charge, followed by transfer to solid support and identification, have been developed. The present compilation brings together a large number of these techniques, some of which are adaptations of the original technique in order to solve a problem, whereas others are far flung and vastly different from how the original techniques were envisioned. The goal of our work is to not only compile the vast array of techniques based on western blot, but also give practical methods. We suspect that almost everyone involved in the enterprise has tried to bring a new technique to their laboratory by reproducing methods found in traditional publications. Doing this is commonly fraught with