Alexander / Block The Separation and Isolation of Proteins

a Laboratory Manual of Analytical Methods of Protein Chemistry (Including Polypeptides)
1. Auflage 2014
ISBN: 978-1-4831-4980-6
Verlag: Elsevier Science & Techn.
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 262 Seiten, Web PDF

A Laboratory Manual of Analytical Methods of Protein Chemistry (Including Polypeptides), Volume 1: The Separation and Isolation of Proteins deals with the techniques used in the separation and isolation of proteins, including fractionation and characterization by dialysis, multi-membrane electrodecantation, and zonal density gradient electrophoresis. The fractionation of proteins by adsorption and ion exchange is also described. This book is comprised of seven chapters and begins with a discussion on procedures for the separation of proteins, paying particular attention to the liberation of proteins from cellular material; removal of lipids from lipoproteins; and denaturation, fractionation, and purification of proteins. The next chapter focuses on the isolation of biologically active proteins such as cytochrome, bacterial amylases, and bacterial proteinases. The reader is methodically introduced to fractionation of proteins by adsorption and ion exchange; fractionation and characterization by dialysis; multi-membrane electrodecantation; and continuous and discontinuous partition. The final chapter explains how zonal density gradient electrophoresis works as a separation method for natural mixtures of proteins, their degradation products, and other substances carrying electric charges in solution or suspension. This volume will be of interest to chemists working with proteins.

Autoren/Hrsg.

Alexander, P.

Block, R. J.

Inhaltsverzeichnis

1;Front Cover
;1
2;The Separation and Isolation of Proteins;4
3;Copyright Page
;5
4;Table of Contents
;6
5;PREFACE;8
6;CONTENTS LIST: VOLUME 2;9
7;CONTENTS LIST: VOLUME 3;9
8;Chapter 1. SEPARATION OF PROTEINS
;11
8.1;A. EXTRACTION OF PROTEINS FROM NATURAL SOURCES;11
8.2;B. REMOVAL OF LIPIDS FROM LIPOPROTEINS;17
8.3;C. DENATURATION OF PROTEINS;17
8.4;D. NEUTRAL SALTS FOR THE FRACTIONATION AND PURIFICATION OF PROTEINS
;18
8.5;E. THE FRACTIONATION AND PURIFICATION OF PROTEINS BY ORGANIC SOLVENTS
;23
8.6;F. THE FRACTIONATION AND PURIFICATION OF PROTEINS WITH HEAVY METAL IONS
;29
8.7;REFERENCES;35
9;Chapter 2. ISOLATION OF BIOLOGICALLY ACTIVE PROTEINS
;41
9.1;CYTOCHROMES OF COW HEART MUSCLE;41
9.2;Cytochromes of Baker's Yeast;51
9.3;CYTOCHROMES OF PSEUDOMONAS AERUGINOSA;62
9.4;BACTERIAL a-AMYLASE
;66
9.5;BACTERIAL PROTEINASE (Hagihara et al, 1958).
;70
9.6;REFERENCES;75
10;Chapter 3. FRACTIONATION OF PROTEINS BY ADSORPTION AND ION EXCHANGE;80
10.1;PART A
;80
10.1.1;A : THE FRACTIONATION AND PURIFICATION OF PROTEINS BY ADSORPTION METHODS

;80
10.1.2;B. THE SEPARATION OF PROTEINS BY COLUMN CHROMATOGRAPHY
;83
10.1.3;REFERENCES;98
10.2;PART B A VARIABLE GRADIENT DEVICE FOR CHROMATOGRAPHY
;101
10.2.1;Simple Gradients;102
10.2.2;Compound Gradients;103
10.2.3;General Characteristics;109
10.2.4;Design and Construction;110
10.2.5;REFERENCES;115
11;Chapter 4. FRACTIONATION AND CHARACTERIZATION BY DIALYSIS
;117
11.1;NATURE OF PROCESS;117
11.2;THE ANALYTICAL DIALYSIS CELL;119
11.3;SEPARATION BY DIALYSIS;124
11.4;THE EFFECT OF TEMPERATURE;126
11.5;SOLVENT EFFECTS;127
11.6;MEMBRANES;129
11.7;MEMBRANE SELECTIVITY;130
11.8;REFERENCES;132
12;Chapter 5. PARTITION
;135
12.1;THE NATURE OF PARTITION;135
12.2;FUNDAMENTAL OPERATION OF THE TRAIN;139
12.3;METHODS OF OPERATION OF A DISTRIBUTION TRAIN;143
12.4;LOADING THE APPARATUS;159
12.5;INTRODUCTION OF THE SAMPLE;162
12.6;ATTAINMENT OF HIGHER PURITY BY SACRIFICING YIELD;163
12.7;TYPES OF COUNTERCURRENT DISTRIBUTION TRAINS AVAILABLE
;165
12.8;CARE AND WASHING OF APPARATUS;170
12.9;ANALYTICAL TECHNIQUES OF SPECIAL VALUE;170
12.10;DETERMINATION OF MOLECULAR WEIGHT BY PARTITION;173
12.11;CONTINUOUS PARTITION;175
12.12;REFERENCES;178
13;Chapter 6. MULTI-MEMBRANE ELECTRODECANTATION
;182
13.1;A. HISTORICAL INTRODUCTION;182
13.2;B. PRINCIPLE AND THEORY;183
13.3;C. CONSTRUCTION OF A MULTI-MEMBRANE ELECTRODECANTATION APPARATUS
;191
13.4;D. OPERATION OF THE MULTI-MEMBRANE ELECTRODECANTATION APPARATUS*
;197
13.5;E. DISCUSSION OF M.M.E.D. AND E-C;204
13.6;F. APPLICATION OF THE METHOD;206
13.7;REFERENCES;211
14;Chapter 7. ZONAL DENSITY GRADIENT ELECTROPHORESIS
;216
14.1;INTRODUCTION;216
14.2;HISTORICAL;217
14.3;STABILITY CONDITIONS;218
14.4;CAPACITY;225
14.5;ELECTROCHEMICAL SEPARATION PRINCIPLES;227
14.6;THE INFLUENCE OF CONDUCTANCE GRADIENTS;229
14.7;THE INFLUENCE OF VISCOSITY GRADIENTS;231
14.8;METHODS FOR PREPARING DENSITY GRADIENT COLUMNS;232
14.9;ELECTRODE REACTION PRODUCTS AND THEIR POSSIBLE INTERFERENCE IN THE SEPARATION
;243
14.10;ELECTROPHORESIS EQUIPMENT AND EXPERIMENTAL PROCEDURE
;246
14.11;METHODS FOR VISUALIZATION OF ZONES IN THE LIQUID COLUMN
;265
14.12;APPLICATIONS AND RESULTS;270
14.13;REFERENCES;277
15;AUTHOR INDEX;280
16;SUBJECT INDEX;285