Physical and Chemical Methods -

Physical and Chemical Methods (eBook)

Methods in Immunology and Immunochemistry, Vol. 2
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2014 | 1. Auflage
480 Seiten
Elsevier Science (Verlag)
978-1-4832-2299-8 (ISBN)
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Methods in Immunology: Volume II, Physical and Chemical Methods is a collection of papers dealing with electrophoresis, analytical ultracentrifugation, dialysis, ultrafiltration, cellulose ion exchangers, and chromatographic separation of macromolecules on porous gels. Some papers explain the applications of radioisotopes, optical analysis, and chemical analysis of proteins, carbohydrates, lipids, and nucleic acid. One paper describes the theory of electro-migration. Factors such as electrical charge or frictional coefficients govern the rate of migration of charged particles in an electric field. The differences found in their velocities can be used to separate substances or analyze them. Mobility is a characteristic property of molecules and can also be influenced by the composition of the medium or solution. Dialysis separates solvents too large to diffuse through a barrier from smaller solutes; ultrafiltration (reverse osmosis) forces solvent and solutes up to a certain critical size through the barrier by a high pressure on one side. The book notes that the membrane never becomes plugged in dialysis because of some opposite movement of the solvent. Another paper points out that the significance of radioactive tracers in immunochemistry employed to identify and label macromolecules functioning as antigens and antibodies. The collection can prove valuable to bio-chemists, cellular biologists, micro-biologists, developmental biologists, and scientists involved in immunological research.
Methods in Immunology: Volume II, Physical and Chemical Methods is a collection of papers dealing with electrophoresis, analytical ultracentrifugation, dialysis, ultrafiltration, cellulose ion exchangers, and chromatographic separation of macromolecules on porous gels. Some papers explain the applications of radioisotopes, optical analysis, and chemical analysis of proteins, carbohydrates, lipids, and nucleic acid. One paper describes the theory of electro-migration. Factors such as electrical charge or frictional coefficients govern the rate of migration of charged particles in an electric field. The differences found in their velocities can be used to separate substances or analyze them. Mobility is a characteristic property of molecules and can also be influenced by the composition of the medium or solution. Dialysis separates solvents too large to diffuse through a barrier from smaller solutes; ultrafiltration (reverse osmosis) forces solvent and solutes up to a certain critical size through the barrier by a high pressure on one side. The book notes that the membrane never becomes plugged in dialysis because of some opposite movement of the solvent. Another paper points out that the significance of radioactive tracers in immunochemistry employed to identify and label macromolecules functioning as antigens and antibodies. The collection can prove valuable to bio-chemists, cellular biologists, micro-biologists, developmental biologists, and scientists involved in immunological research.

Front Cover 1
Physical and Chemical Methods 4
Copyright Page 5
Table of Contents 10
Contributors to Volume II 6
Contents of Other Volumes 16
Chapter 6. Electrophoresis 22
A. 

22 
B. Moving-Boundary Electrophoresis 27
C. Analytical Zone Electrophoresis 40
D. Preparative Zone Electrophoresis 78
Chapter 7. Preparative and Analytical Ultracentrifugation 102
A. Introduction 102
B. Moving-Boundary Methods 110
C. Moving-Zone Methods 120
D. Isodensity Methods 126
E. Separation of Subcellular Particles 129
F. Classical Sedimentation Equilibrium Methods 133
G. Summary of Mathematical Formulas 134
Chapter 8. Dialysis and Ultrafiltration 140
A. Introduction 140
B. The Membrane 140
C. Apparatus 142
D. Methods for Altering Porosity 152
Chapter 9. Chromatographic Separations of Macromolecules on Porous Gels and Cellulose Ion Exchangers 156
A. Introduction 156
B. Chromatography on Gels 156
C. Cellulose Ion Exchange Chromatography 175
D. Analysis of Column Effluents 181
E. Recovery of Proteins 182
Chapter 10. Optical Analysis 184
A. Applications of Ultraviolet and Visible Spectroscopy 184
B. Turbidimetric Assay Methods: Application to 
195 
Chapter 11. Radioisotopes and Their Applications 204
A. Labeling of Macromolecules 204
B. Principles in Choice of Radioactive Tracers 206
C. The Radioisotope Laboratory and Handling Procedures 215
D. Analytical Equipment 220
E. Sample Preparation 228
F. Evaluation of Counting Data 239
G. Autoradiography in Immunological Investigation 249
H. Glossary of Symbols, Abbreviations, and Terms 261
Chapter 12. Chemical Analysis 270
A. Protein Analysis 270
B. Carbohydrate Analysis 303
C. Preparation of Lipoproteins and Analysis of Lipids 326
D. Nucleic Acid Analysis 338
APPENDIX 
364 
APPENDIX 
386 
1. General Remarks 386
2. The Formulation of Buffers 388
3. Use of Reference Standard Buffer Solutions 389
4. Sodium Hydroxide for Use in Buffers 391
5. Hydrochloric Acid for Use in Buffers 392
6. Chemicals for Preparation of Buffers 392
7. Various Buffer Systems, with References 402
8. Composition of Selected Buffers 407
9. Indicators 426
10. Isotonic Solutions 428
References 428
APPENDIX 
430 
AUTHOR INDEX 436
SUBJECT INDEX 447

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