Basic Protein and Peptide Protocols / Edition 1

by John M. Walker

Overview

Basic Protein and Peptide Prools offers an excellent collection of reproducible, step-by-step laboratory methods covering three major areas: (1) the quantitation and characterization of proteins, (2) the electrophoretic and blotting procedures used in protein isolation and characterization, and (3) the analysis of protein and peptide structure. THOUSANDS of labs are already using Basic Protein and Peptide Prools-you should be too!

Product Details

ISBN-13:	9780896032682
Publisher:	Springer-Verlag New York, LLC
Publication date:	04/26/1994
Series:	Methods in Molecular Biology , #32
Edition description:	1994
Pages:	490
Product dimensions:	6.10(w) x 9.25(h) x 0.04(d)

About the Author

Walker, John M. (Univ of Hertfordshire)

The contributors represent the specialties of biology, chemistry, cancer research and mollecular oncology. They come from the U.K., U.S., Australia, Germany, Sweeden and Northern Ireland. Institutions prominently represented include Kings Coll-London, Heart Science Center, and the Queen's Univ in Belfast.

Table of Contents

The Lowry Method for Protein Quantitation.- The Bicinchoninic Acid (BCA) Assay for Protein Quantitation.- The Bradford Method for Protein Quantitation.- Nondenaturing Polyacrylamide Gel Electrophoresis of Proteins.- SDS Polyacrylamide Gel Electrophoresis of Proteins.- Gradient SDS Polyacrylamide Gel Electrophoresis of Proteins.- Acetic Acid-Urea Polyacrylamide Gel Electrophoresis of Proteins.- Electrophoresis of Peptides.- Isoelectric Focusing of Proteins in Ultra-Thin Polyacrylamide Gels.- The In Vivo Isotopic Labeling of Proteins for Polyacrylamide Gel Electrophoresis.- Two-Dimensional Polyacrylamide Gel Electrophoresis of Proteins.- Two-Dimensional Polyacrylamide Gel Electrophoresis Using Immobilized pH Gradients in the First Dimension.- Two-Dimensional Polyacrylamide Gel Electrophoresis Using Flat-Bed Isoelectric Focusing in the First Dimension.- Quantification of Proteins on Polyacrylamide Gels (Nonradioactive).- Detection of Proteins in Polyacrylamide Gels Using an Ultrasensitive Silver Staining Technique.- Identification of Glycoproteins on Nitrocellulose Membranes and Gels.- The Release of Oligosaccharides from Glycoproteins.- Structural Profiling of Oligosaccharides of Glycoproteins.- Drying Gels.- Fluorography of Polyacrylamide Gels Containing Tritium.- The Electrophoretic Elution of Proteins from Polyacrylamide Gels.- Preparation of Proteins from Gels for Protein Microsequencing.- Comparison of Protein Primary Structures.- Protein Blotting.- Detection of Polypeptides on Immunoblots Using Secondary Antibodies or Protein A.- Detection of Proteins on Blots Using the Avidin-Biotin System.- Detection of Proteins on Protein Blots Using Chemiluminescent Systems.- The Detection of Proteins on Blots Using Gold or Immunogold.- Amino Acid Analysis.- Molecular-Weight Estimation for Native Proteins Using Size-Exclusion High-Performance Liquid Chromatography.- Peptide Purification by Reverse-Phase HPLC.- Enzymatic Methods for Cleaving Proteins.- Chemical Cleavage of Proteins.- Chemical Modification of Proteins.- The Dansyl Method for Identifying N-Terminal Amino Acids.- The Dansyl-Edman Method for Peptide Sequencing.- C-Terminal Sequencing of Peptides.- Analysis of Cysteine Residues and Disulfide Bonds.- Preparation and Characterization of Monoclonal Antibodies to Proteins and Other Cellular Components.- The Raising of a Polyclonal Antiserum to a Protein.- Production of Antisera to Synthetic Peptides.- Production of Antibodies Using Proteins in Gel Bands.- Preparation of Purified Immunoglobulin G (IgG).- Enzyme and Biotin Labeling of Antibody.- Labeling of Peptides and Proteins by Radioiodination.- Radioimmunoassay of Peptides and Proteins.- Enzyme-Linked Immunosorbent Assay (ELISA).- Free Zone Capillary Electrophoresis.

Editorial Reviews

...would I buy this volume? I think that the answer to this is yes. It is a useful and concise volume that would be of great value around the lab. I would also recommend it for libraries as it provides an excellent reference source on techniques of protein analysis for undergraduates entering the difficult world of research projects. The value of this volume lies in the fact that any potential reader is liable to use the techniques described in several chapters to pursue a piece of research work. - Trends in Biochemical Sciences

From the Publisher

3 Stars from Doody

This book is written to update an earlier volume in the Methods in Molecular Biology series and is aimed at researchers who have become involved in the biochemical analyses of proteins and peptides. The text contains both newly developed protocols as well as revised established procedures using new technologies. The analytical methods included here reflect newer technologies such as capillary electrophoresis, immobilized pH gradients in isoelectric focusing, immunogold reagents and the BCA assay for proteins. Also included are newer applications that improve sensitivities, resolution, speed, or ease of use, including chemiluminescent systems and lectins. Each protocol provides an introduction, list of materials needed, step-by-step methods, notes, and references. This book is written as a bench top manual and guide for those new to protein biochemistry. A researcher not familiar with the procedure should be able to follow the directions without difficulty. The figures that are included clearly illustrate the types of results and sensitivities to be expected with the protocols. The references listed cover the development of the technique as published in the literature. The table of contents and index are adequate. This book will serve as an excellent guidebook for researchers who are trained as molecular biologists and have become interested in the analytical methods of biochemistry. Each chapter lists advantages and disadvantages including sensitivity and limitations. This is an excellent bench top edition.

Ada A. Cole

An update to Walker's classic Proteins (1984), adding many new tested basic laboratory methods for protein and peptide analysis, for both those new to the protein chemistry laboratory and more established workers who wish to try a new technique for the first time. Among the key protocols included are protein and peptide electrophoresis techniques, gel staining methods, methods for protein quantitation, peptide mapping, protein blotting and various detection systems, peptide sequencing, glycoprotein analysis, production of polyclonal and monoclonal antibodies, ELISA, the use of immobilized pH gradients in isoelectric focusing, IgG purification, the use of immunogold reagents, and the detection and analysis of thiol groups. Plastic comb binding. Annotation c. Book News, Inc., Portland, OR (booknews.com)

Booknews

Reviewer: Ada A. Cole, PhD (Rush Medical College of Rush University)
Description: This book is written to update an earlier volume in the Methods in Molecular Biology series and is aimed at researchers who have become involved in the biochemical analyses of proteins and peptides. The text contains both newly developed protocols as well as revised established procedures using new technologies.
Purpose: The analytical methods included here reflect newer technologies such as capillary electrophoresis, immobilized pH gradients in isoelectric focusing, immunogold reagents and the BCA assay for proteins. Also included are newer applications that improve sensitivities, resolution, speed, or ease of use, including chemiluminescent systems and lectins. Each protocol provides an introduction, list of materials needed, step-by-step methods, notes, and references.
Audience: This book is written as a bench top manual and guide for those new to protein biochemistry. A researcher not familiar with the procedure should be able to follow the directions without difficulty.
Features: The figures that are included clearly illustrate the types of results and sensitivities to be expected with the protocols. The references listed cover the development of the technique as published in the literature. The table of contents and index are adequate.
Assessment: This book will serve as an excellent guidebook for researchers who are trained as molecular biologists and have become interested in the analytical methods of biochemistry. Each chapter lists advantages and disadvantages including sensitivity and limitations. This is an excellent bench top edition.

Doody's Review Service