상세정보

New textbooks at all levels of chemistry appear with great regularity. Some fields such as basic biochemistry, organic re action 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 real lack of up-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 that is advancing the field. It is not often easy to persuade such individuals to set time aside to help spread the knowledge they have accumulated. 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 instruc­ tive introductions to their fields. These should serve the needs of one semester or one quarter graduate courses in chemistry and biochemistry. In so me cases the availability of texts in active research areas should help stimulate the creation of new courses.

New textbooks at all levels of chemistry appear with great regularity. Some fields such as basic biochemistry, organic re action 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 real lack of up-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 that is advancing the field. It is not often easy to persuade such individuals to set time aside to help spread the knowledge they have accumulated. 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 instruc­ tive introductions to their fields. These should serve the needs of one semester or one quarter graduate courses in chemistry and biochemistry. In so me cases the availability of texts in active research areas should help stimulate the creation of new courses.

정보제공 :

CONTENTS
Series Preface = ⅴ
Preface to the Third Edition = ⅶ
Preface to the Second Edition = ⅸ
Preface to the First Edition = xi
Chapter 1 The Protein Purification Laboratory = 1
  1.1 Apparatus, Special Materials, and Reagents = 1
  1.2 Separation of Precipitates and Particulate Material = 3
    Filtration = 3
    Centrifugation = 4
  1.3 Principles of Column Chromatography = 8
  1.4 Manipulation of Protein Solutions = 14
    Concentration = 15
    Removal of Salts； Changing Buffers = 17
Chapter 2 Making an Extract = 22
  2.1 The Raw Material = 22
    Freshness and Storage = 25
  2.2 Cell Disintegration and Extraction = 26
    Mammalian Tissues = 30
    Erythrocytes = 31
    Soft Plant Tissues = 31
    Yeasts = 31
    Bacteria = 32
    Fatty Tissues = 34
  2.3 Optimization and Clarification of the Extract = 34
  2.4 Extraction of Membrane Proteins = 38
Chapter 3 Analysis―Measurement of Protein and Enzyme Activity = 44
  3.1 Methods for Measuring Protein Concentration = 44
    Biuret Reaction = 45
    Lowry Method = 46
    UV Absorption = 46
    Dye Binding = 48
    Bicinchonic Acid = 48
  3.2 Measurement of Enzyme Activity―Basic Principles = 50
    Substrate Concentration, Activators, and Inhibitors = 50
    pH, Ionic Strength, and Temperature = 55
  3.3 Measurement of Enzyme Activity Using Stopped Methods = 56
    Incubation Conditions = 57
    Stopping Methods = 58
    Measurement of Product = 59
  3.4 Measurement of Enzyme Activity Using Continuous Methods = 62
    Coupled Methods = 63
  3.5 Practical Points in Enzyme Activity Determination = 68
Chapter 4 Separation by Precipitation = 71
  4.1 General Observations = 71
  4.2 The Solubility of Proteins at Low Salt Concentrations = 72
    Points to Note in Practice = 75
  4.3 Salting Out at High Salt Concentration = 76
  4.4 Precipitation with Organic Solvents = 85
    General Theory = 85
    Choice ot Solvent = 87
    Operating Procedures = 89
  4.5 Precipitation with Organic Polymers and Other Materials = 92
  4.6 Affinity Precipitation = 93
  4.7 Precipitation by Selective Denaturation = 95
    General Principles = 95
    Temperature Denaturation = 96
    pH Denaturation = 98
    Denaturation by Organic Solvents = 100
Chapter 5 Separation by Adsorption I: General Principles = 102
  5.1 General Chromatographic Theory = 103
    Partition Coefficients = 103
    Zone Spreading, Resolution, and the Plate Height Concept = 105
    The Dissociation Constant for Protein-Adsorbent Interaction = 111
    Simplified Theory of Adsorption = 112
  5.2 Membrane Adsorbents； Radial Flow Columns = 119
  5.3 Batch Adsorption = 121
    General Principles = 121
    Practical Approaches = 123
  5.4 High-Performance Liquid Chromatography = 126
    General Principles = 126
    Relationships Between Bead Size, Flow Rate, Pressure, and Optimum Performance = 128
  5.5 Types of Adsorbent Used in Protein Chromatography = 132
    Nature of the Bead Matrix = 132
    Summary of Adsorbent Types = 135
  5.6 Operating Conditions for Column Chromatography = 139
    Sample Application = 139
    Overload and Displacement Chromatography = 139
    Flow Rates = 142
Chapter 6 Separation by Adsorption II: Ion Exchangers and Nonspecific Adsorbents = 146
  6.1 Ion Exchangers―Principles, Properties, and Uses = 146
    General Principles = 146
    Adsorptive Capacities of Ion Exchangers = 150
    Types of Ion Exchangers = 152
    pH and Donnan Effects = 153
    Elution of Adsorbed Protein = 154
  6.2 Ion-Exchange Chromatograpahy―Practical Aspects = 157
    Trials to Determine Ion-Exchange Behavior = 159
    Buffers for Use in Ion-Exchange Chromatography = 160
    Conditions of Adsorption = 164
    Size and Dimensions of the Column = 165
    Procedures for Elution = 167
  6.3 Inorganic Adsorbents = 172
    Hydroxyapatite and Calcium Phosphate Gels = 173
  6.4 Hydrophobic Adsorbents = 175
    Application of Sample to a Hydrophobic Column = 176
    Elution of Protein from Hydrophobic Columns = 177
    Reverse Phase Chromatography = 178
    Other Hydrophobic Techniques = 179
  6.5 Immobilized Metal Affinity Chromatography (IMAC) = 180
    General Principles = 180
    Operating Conditions for IMAC = 182
  6.6 Miscellaneous Adsorbents = 183
    Cationic Polymer-Nucleic Acid Complexes as Batch Adsorbents = 183
    Thiophilic Adsorbents = 184
    Mixed-Function Adsorbents = 185
Chapter 7 Separation by Adsorption―Affinity Techniques = 187
  7.1 Principles of Affinity Chromatography = 187
    Synthesis of Affinity Adsorbents = 188
    Application of Chromatographic Theory to Affinity Adsorbents = 196
    General Techniques and Procedures in Affinity Adsorption Chromatography = 200
  7.2 Immunoadsorbents = 204
    Basic Principles = 205
    Methods Using Polyclonal Antibodies = 205
    Methods Using Monoclonal Antibodies = 208
    Relative Advantages of Polyclonal and Monoclonal Antibodies = 209
  7.3 Dye Ligand Chromatography = 210
    Developmental History = 210
    Preparation of Dye Ligand Adsorbents = 214
    Dye-Protein Interactions = 217
    Screening to Obtain a Suitable Adsorbent = 219
    Elution of Proteins and Enzymes from Dye Columns = 223
    Cleaning and Storage of Dye Adsorbents = 224
  7.4 Affinity Elution from Ion Exchangers and Other Adsorbents = 226
    Affinity Elution from Ion Exchangers = 227
    Affinity Elution from Other Adsorbents = 231
    Practice and Theory of Affinity Elution = 233
  7.5 Commonly Used Affinity and Pseudo-Affinity Adsorbents = 236
    Small Ligands = 236
    Biopotymer Ligands = 236
Chapter 8 Separation in Solution = 238
  8.1 Gel Filtration = 238
    Practical Procedures = 243
  8.2 Electrophoretic Methods = 250
    Electrophoresis Principles = 251
    Methods for Preparative Electrophoresis―Horizontal Slabs = 253
    Methods for Preparative Eleclrophoresis―Vertical Systems = 255
    Buffer Systems for Electrophoresis = 256
    Isoetectric Focusing = 258
    Isotachophoresis = 262
  8.3 Liquid Phase Partitioning = 264
  8.4 Ultrafiltration = 267
Chapter 9 Purification of Special Types of Proteins = 270
  9.1 Recombinant Proteins = 270
    Terminology of Recombinant Proteins = 271
  9.2 Membrane Proteins = 277
  9.3 Purification of Antibodies = 279
Chapter 10 Small-Scale and Large-Scale Procedures = 283
  10.1 Small-Scale Procedures―Proteins for Sequencing = 283
  10.2 Large-Scale Procedures = 287
    Scaling Up in the Laboratory = 287
    Commercial-Scale Protein Production = 291
Chapter 11 Analysis for Purity = 293
  11.1 Electrophoretic Analysis = 293
    Simple (Native) Gel Electrophoresis = 294
    Urea Gels = 296
    SDS Gels = 296
    Gradieni Gels = 298
    Isoelectric Focusing = 299
    Two-Dimensional Systems = 300
    Capillary Electrophoresis = 300
    Staining and Detection of Proteins after Electrophoresis = 302
    Detection of Specific Proteins = 303
  11.2 Other Analytical Methods = 307
Chapter 12 Optimization of Procedures；Final Steps = 310
  12.1 Speed Versus Resolution: The Time Factor = 311
  12.2 Stabilizing Factors for Enzymes and Other Proteins = 317
    Prevention of Denaturation = 317
    Avoidance of Catalytic Site Inaclivation = 318
    Avoidance of Proteolytic Degradation = 321
    Other Stabilizing Influences on Proteins = 323
  12.3 Control of pH: Buffers = 324
    Buffer Theory = 324
    Effect of Temperature, Ionic Strength, and Organic Solvents on pK pKa Values = 326
    Making Up Buffer Solutions = 330
  12.4 Following a Published Procedure = 333
  12.5 Final Steps―Storage, Crystallization, and Publication = 335
    Crystallization for Purification = 336
    Methods for Crystallization for X-ray Diffraction Studies = 337
    Conditions for Storage of Purified Proteins = 342
    What is Important for Publication? = 344
Appendix A Precipitation Tables = 346
Appendix B Solutions for Measuring Protein Concentration = 349
Appendix C Buffers for Use in Protein Chemistry = 351
Appendix D Chromatographic Materials = 353
References = 356
Index = 375