Protein Analysis And Purification

This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.

How one goes about analyzing proteins is a constantly evolving ?eld that is no longer solely the domain of the protein biochemist. Inves- gators from diverse disciplines ?nd themselves with the unanticipated task of identifying and analyzing a protein and studying its physical properties and biochemical interactions. In most cases, the ultimate goal remains understanding the role(s) that the target protein is playing in cellular physiology. It was my intention that this manual would make the initial steps in the discovery process less time consuming and less intimidating. This book is not meant to be read from cover to cover. The expanded Table of Contents and the index should help locate what you are seeking. My aim was to provide practically oriented information that will assist the experimentalist in benchtop problem solving. The appendices are ?lled with diverse information gleaned from catalogs, handbooks, and manuals that are presented in a distilled fashion designed to save trips to the library and calls to technical service representatives. The user is encouraged to expand on the tables and charts to ?t individual experimental situations. This second edition pays homage to the computer explosion and the various genome projects that have revolutionized how benchtop scienti?c research is performed. Bioinformatics and In silico science are here to stay. However, the second edition still includes recipes for preparing buffers and methods for lysing cells.

New textbooks at alllevels of chemistry appear with great regularity. Some fields like basic biochemistry, organic reaction mechanisms, and chemical thermodynamics are weil represented by many excellent texts, and new or revised editions are published sufficiently often to keep up with progress in research. However, some areas of chemistry, especially many of those taught at the graduate level, suffer from a reallack ofup-to-date textbooks. The most serious needs occur in fields that are rapidly changing. Textbooks in these subjects usually have to be written by scientists actually involved in the research which is advancing the field. lt is not often easy to persuade such individuals to settime aside to help spread the knowledge they have accumu lated. Our goal, in this series, is to pinpoint areas of chemistry where recent progress has outpaced what is covered in any available textbooks, and then seek out and persuade experts in these fields to produce relatively concise but instructive introductions to their fields. These should serve the needs of one semester or one quarter graduate courses in chemistry and biochemistry. In some cases the availability of texts in active research areas should help stimulate the creation of new courses. NewYork CHARLES R. CANTOR Preface to the Second Edition The original plan for the first edition of this book was to title it Enzyme Purification: Princip/es and Practice.

The methods found here are drawn from such popular laboratory manuals as "Proteins and Proteomics" and "Purifying Proteins for Proteomics." This volume contains an essential collection of purification methods using gel electrophoresis and column chromatography.

This is a state-of-the-art sourcebook on modern high-resolution biochemical separation techniques for proteins. It contains all the basic theory and principles used in protein chromatography and electrophoresis.

Guide to Protein Purification , designed to serve the needs of the student, experienced researcher and newcomer to the field, is a comprehensive manual that provides all the up-to-date procedures necessary for purifying, characterizing, and handling proteins and enzymes in one source. Key Features* Detailed procedures newly written for this volume* Extensive practical information* Rationale and strategies for protein and enzyme purification* Personal perspectives on enzyme purification by eminent researchersAmong the Topics Covered* General methods for handling proteins and enzymes * Extraction, subcellular fractionation, and solubilization procedures * Comprehensive purification techniques * Specialized purification procedures * Protein characterization * Immunological procedures * Computer analysis of protein structure.