From Reviews of the First Edition
"...a must for researchers in protein chemistry and especially those with less experience..."
-Doody's Health Sciences Book Review Journal
"...definitely an indispensable benchtop handbook...highly recommended for all levels of research
within the field."-Bioseparation
"...a very useful tool for daily work." -Nahrung-Food

Since publication of the bestselling first edition of John Walker's widely acclaimed Protein Protocols Handbook in 1996, there have been many highly significant developments in protein biochemistry. This greatly enhanced second edition introduces 60 critically important new chapters dealing with topics and techniques that have emerged since the first edition, as well as significantly updating the remaining articles. The new chapters cover many rapidly developing areas, particularly the application of mass spectrometry in protein characterization, as well as the now well-established 2-D PAGE technique in proteomics. The section on glycoprotein analysis has also been significantly expanded, and methods for the production of single-chain and phage-displayed antibodies have been added to the section on antibodies.
Each readily reproducible method follows the highly praised format of the Methods in Molecular Biology(TM) series, offering a concise summary of its basic theory, a complete materials list, a step-by-step protocol for its successful execution, and extensive notes on avoiding pitfalls, or on modifying the method to function within your own experimental circumstances. The authors are commonly the techniques' originators, and each has demonstrated a hands-on mastery of the methods described, always fine-tuning them here for optimal productivity.
Comprehensive, cutting-edge, and highly practical, The Protein Protocols Handbook, Second Edition is today's indispensable benchtop manual and guide-not only for all those new to the protein chemistry laboratory, but also for those established workers seeking to broaden their armametarium of techniques in the urgent search for rapid and still more robust results.

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 Biology(TM) 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.

Part I. Quantitation of Proteins 1 Protein Determination by UV Absorption Alastair Aitken and Michèle P. Learmonth 2 The Lowry Method for Protein Quantitation Jakob H. Waterborg 3 The Bicinchoninic Acid (BCA) Assay for Protein Quantitation John M. Walker 4 The Bradford Method for Protein Quantitation Nicholas J. Kruger 5 Ultrafast Protein Determinations Using Microwave Enhancement Robert E. Akins and Rocky S. Tuan 6 The Nitric Acid Method for Protein Estimation in Biological Samples Scott A. Boerner, Yean Kit Lee, Scott H. Kaufmann, and Keith C. Bible 7 Quantitation of Tryptophan in Proteins Alastair Aitken and Michèle P. Learmonth 8 Flow Cytometric Quantitation of Cellular Proteins Thomas D. Friedrich, F. Andrew Ray, Judith A. Laffin, and John M. Lehman 9 Kinetic Silver Staining of Proteins Douglas D. Root and Kuan Wang Part II. Electrophoresis of Proteins and Peptides and Detection in Gels 10 Nondenaturing Polyacrylamide Gel Electrophoresis of Proteins John M. Walker 11 SDS Polyacrylamide Gel Electrophoresis of Proteins John M. Walker 12 Gradient SDS Polyacrylamide Gel Electrophoresis of Proteins John M. Walker 13 SDS-Polyacrylamide Gel Electrophoresis of Peptides Ralph C. Judd 14 Identification of Nucleic Acid Binding Proteins Using Nondenaturing Sodium Decyl Sulfate Polyacrylamide Gel Electrophoresis (SDecS-Page) Robert E. Akins and Rocky S. Tuan 15 Cetyltrimethylammonium Bromide Discontinuous Gel Electrophoresis of Proteins: Mr-Based Separation of Proteins with Retained Native Activity Robert E. Akins and Rocky S. Tuan 16 Acetic-Acid-Urea Polyacrylamide Gel Electrophoresis of Basic Proteins Jakob H. Waterborg 17 Acid-Urea-Triton Polyacrylamide Gel Electrophoresis of Histones Jakob H. Waterborg 18 Isoelectric Focusing of Proteins in Ultra-Thin Polyacrylamide Gels John M.Walker 19 Protein Solubility in Two-Dimensional Electrophoresis: Basic Principles and Issues Thierry Rabilloud 20 Preparation of Protein Samples from Mouse and Human Tissues for 2-D Electrophoresis Joachim Klose 21 Radiolabeling of Eukaryotic Cells and Subsequent Preparation for 2-D Electrophoresis Nick Bizios 22 Two-Dimensional Polyacrylamide Gel Electrophoresis Using Carrier Ampholyte pH Gradients in the First Dimension Patricia Gravel 23 Casting Immobilized pH Gradients (IPGs) Elisabetta Gianazza 24 Nonequilibrium pH Gel Electrophoresis (NEPHGE) Mary F. Lopez 25 Difference Gel Electrophoresis Mustafa Ünlü and Jonathan Minden 26 Comparing 2-D Electrophoretic Gels Across Internet Databases Peter F. Lemkin and Gregory C. Thornwall 27 Immunoblotting of 2-D Electrophoresis Separated Proteins Barbara Magi, Luca Bini, Sabrina Liberatori, Roberto Raggiaschi, and Vitaliano Pallini 28 Quantification of Radiolabeled Proteins in Polyacrylamide Gels Wayne R. Springer 29 Quantification of Proteins on Polyacrylamide Gels Bryan John Smith 30 Rapid and Sensitive Staining of Unfixed Proteins in Polyacrylamide Gels with Nile Red Joan-Ramon Daban, Salvador Bartolomé, Antonio Bermúdez, and F. Javier Alba 31 Zinc-Reverse Staining Technique Carlos Fernandez-Patron 32 Protein Staining with Calconcarboxylic Acid in Polyacrylamide Gels Jung-Kap Choi, Hee-Youn Hong, and Gyurng-Soo Yoo 33 Detection of Proteins in Polyacrylamide Gels by Silver Staining Michael J. Dunn 34 Protein Detection on Polyacrylamide Gels and Electroblots Using Transition Metal Chelate Stains Wayne F. Patton 35 Detection of Proteins in Polyacrylamide Gels by Fluorescent Staining Michael J. Dunn 36 Detection of Proteins and Sialoglycoproteins in Polyacrylamide Gels Using Eosin Y Stain Fan Lin and Gary E. Wise 37 Electroelution of