Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis: with 16 tables
Saved in:
| Main Author: | |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
Berlin [u.a.]
de Gruyter
1971
|
| Edition: | 2., rev. and expanded ed. |
| Series: | Working methods in modern science
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | XVI, 222 S. Ill., graph. Darst. |
| ISBN: | 3110034956 |
Staff View
MARC
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| 245 | 1 | 0 | |a Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis |b with 16 tables |c by H. Rainer Maurer |
| 250 | |a 2., rev. and expanded ed. | ||
| 264 | 1 | |a Berlin [u.a.] |b de Gruyter |c 1971 | |
| 300 | |a XVI, 222 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 0 | |a Working methods in modern science | |
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Record in the Search Index
| _version_ | 1804115367058997248 |
|---|---|
| adam_text | Disc Electrophoresis
and Related Techniques of
Polyacrylamide Gel
Electrophoresis
by H Rainer Maurer
Dr rer „*
Max-Planck-Institut fur Virus-Forschung
Tubingen, Germany
Second revised and expanded edition
With 88 figures, 16 tables, 948 literature references
w
DE
G Walter de Gruyter • Berlin New York 1971
Table of Contents
Preface v
Acknowledgements ix
1 Theoretical Background 1
1 1 The Polyacrylamide Gel 1
111 Formation and Structure of the Gel 1
112 Polymerization: Catalysts and Processes 3
113 The Molecular Sieving Effect in Polyacrylamide Gel
By W Thorun and H R Maurer 4
1131 Introduction 4
1132 Relations between Gel Density (Pore Size), Cross-
Linkage, Particle Size and Electrophoretic
Migration 5
114 Molecular Weight Determination by Polyacrylamide
Gel Electrophoresis 8
1141 Proteins By W Thorun and H R Maurer 8
1142 Nucleic Acids 17
115 Discrimination of Proteins Similar or Different in Charge
and/or Size by Polyacrylamide Gel Electrophoresis 19
1 2 Important Principles of Electrophoresis 20
1 3 Theory of Disc Electrophoresis: Polyacrylamide Gel Electro-
phoresis with Discontinuous Voltage and PH Gradients
(Ornstein-Davis-System) 22
131 Introduction and General Comments 22
1311 Concentrating Effect 22
1312 Advantages and Disadvantages~of~Continuous
and Discontinuous Buffer and pH Systems 24
132 Processes during Disc Electrophoresis 25
1321 Concentration of the Sample Solution 25
1322 Separation of Sample Components 28
133 Guidelines for the Choice of Suitable Buffer and pH
Systems 29
1 4 Polyacrylamide Gel Electrophoresis with Discontinuous Voltage
Gradients (Allen-System) 29
xii Table of Contents
2 Methods of Analytical Disc Electrophoresis 32
2 1 Physicochemical Properties of the Gel Components and their
Identification 32
211 General Comments 32
212 Concentration Determinations 32
2121 Determination of Acrylamide and N,N -
methylene-bis-acrylamide 33
213 Purity and Purification Methods 34
214 Toxicity of Acrylamide 35
2 2 Equipment for Disc Electrophoresis 35
221 Gel Tubes and Tube Holders 35
222 Apparatus and Accessoires 36
223 Electrophoretic Destaining Equipment 39
224 Power Supply Units 40
2 3 Procedures 41
231 Gel Systems and their Compositions 41
2311 General Comments 41
2312 Explanations to Table 4 43
232 Preparation of Cylindrical Gels 52
2321 Preparation of the Spacer Gel 52
2322 Preparation of the Separation Gel 52
2323 Alternative Gel Preparation 53
233 Problems of the Preparation of Spacer and Separation Gels 54
2331 Effects of Catalysts and Gel Components on
Gel Polymerization, Gelling Time and Electro-
phoretic Migration 54
2332 Water Layering 56
2333 High Density Separation Gels 56
2334 Sources of Error in Faulty Separations 56
234 Modifications of Gel Compositions and Suggestions for
Special Separation Problems (Modifications of Disc
Electrophoresis) 57
2341 Sample, Spacer and Separation Gel: Sequence of
Preparation 57
2342 Band Curvature and Temperature Effects 57
2343 The Use of the Sample Gel 58
2344 Modification of the Spacer Gel 58
H2345 Suspension of the Spacer Gel and Pre-Electro-
phoresis ^ 5 9
2346 Persulfate and Irradiation Artifacts 59
2347 Solubilizable Polyacrylamide Gels 60
2348 Gradient Gel Electrophoresis 60
2349 Two-Dimensional Polyacrylamide Gel Electro-
phoresis 63
234 10 Two-Directional (Double) Disc Electro-
phoresis 64
Table of Contents xiii
234 11 Sephadex Sandwich Disc Electrophoresis 65
234 12 Split Gel Technique 65
234 13 Disc Re-Electrophoresis 65
234 14 PH-Reversal Method 66
234 15 Agarose-Polyacrylamide Composite Gels 66
234 16 Gel Electrophoresis with Detergents 67
234 17 Disc Electrophoresis of Fluorescent-labeled
Proteins 67
234 18 Disc Tissue Electrophoresis 68
234 19 Detection of Biological Activity of Electro-
phoresed Proteins by Tissue Culture on Gels 68
235 Preparation of Sample Solution 68
236 Measuring of the Sample Solution 69
237 Electrophoretic Procedure 69
238 Detection of Separated Substances by Staining 71
2381 Removal of Gels from Gel Tubes 71
2382 Detection Reagents and Color Reactions 72
23821 General Color Reactions of Proteins 72
23822 Color Reactions of Metal-containing
Proteins 76
23823 Color Reactions of Lipoproteins 76
23824 Color Reactions of Glycoproteins and
Mucopolysaccharides 76
23825 Color Reactions of Specific Proteins 78
23826 Color Reactions of Deoxyribonucleic
Acid (DNA) 78
23827 Color Reactions of Ribonucleic
Acids (RNA) 79
2383 Staining 80
2384 Destaining 80
2385 Comments on Staining and Destaining 82
239 Quantitative Evaluation 82
2391 Direct Densitometric Analysis 82
23911 Scanning of Stained Patterns and
Problems of Quantitative Den-
sitometry 83
23912 Recording of Unstained Patterns by
Direct Ultraviolet Absorbance
-Measurement 84
23913 Special Densitometers 86
2392 Photographic-densitometric-Analysis 88
2 3 10 Slicing of the Gel 90
2 3 11 Detection of Radioactive Substances 93
2 3 11 1 Radioactivity Assays in Gel Sections 94
2 3 11 1 1 Direct Counting 94
2 3 11 1 2 Elution Method 95
2 3 11 1 3 Combustion Methods 95
2 3 11 1 4 Hydrolysis Method 96
2 3 11 1 5 Free Radical Depolymerization
Method 98
2 3 11 2 Autoradiography of Longitudinal Gel Sections 99
xiv Table of Contents
2 3 12 Drying of Polyacrylamide Gels for Storage or Radio-
activity Determination 101
2 4 Disc Enzyme Electrophoresis 102
241 Elution Method 103
242 Gel Staining Method 103
243 Substrate and Primer Inclusion Techniques 103
244 Contact Print Method 106
2 5 Disc Immunoelectrophoresis 106
251 Two-dimensional Disc Immunoelectrophoresis 106
2511 Immunodiffuskm in Agar Gel 106
25111 Plain Agar Method 107
25112 Antibody-Agar Method 107
25113 Agar Embedding Method 108
2512 Immunodiffusion in Polyacrylamide Gel 109
252 Unidimensional Disc Immunoelectrophoresis 109
3 Micro-Disc Electrophoresis I l l
3 1 General Methods I l l
3 2 Micro-Disc Electrophoresis combined with Micro-Enzyme and
Immunoanalysis 114
4 Preparative Polyacrylamide Gel Electrophoresis 117
4 1 General Comments 117
411 Gel Preparation 118
412 Sample Loading 118
413 Artifacts mainly arising in Preparative Disc Electro-
phoresis 118
4 2 Methods of Preparative Polyacrylamide Gel Electrophoresis 119
421 Extraction of Gel Fractions 119
4211 Elution by Diffusion 119
4212 Elution by Electrophoresis 119
422 Continuous Separation and Elution by Electrophoresis 120
4221 Micropreparative Devices ^— 120
4222 Preparative Apparatus for Milligram Sample
Quantities 121
423 Discontinuous Separation and Elution by Electrophoresis 130
5 Gel Isoelectric Focusing 132
5 1 General Methods 132
5 2 Procedures and Instructions 133
521 Sample Application 133
Table of Contents xv
522 Staining and Destaining Procedures 136
523 General Comments 136
6 Applications in Clinical Chemistry 137
6 1 Blood Proteins 137
611 Serum Proteins 137
6111 General Remarks 137
6112 Normal Sera 138
6113 Pathological Sera 142
6114 Immunoglobins, Antibodies and Complements 146
6115 Serum Enzymes 147
61151 General Remarks 147
61152 Reactions for the Detection of some
Enzymes 148
612 Blood-Clotting and Platelet Proteins 150
613 Hemoglobin Determination 151
614 Ceruloplasmin 151
615 Nonhemoglobin Proteins of Erythrpcytes 151
6 2 Proteins of other Body Fluids 152
621 Cerebrospinal Fluid (CSF) 152
622 Urine 153
623 Saliva 154
624 Exudates, Transudates and other Body Fluids 154
6 3 Tissue Enzymes 155
6 4 Other Tissue Proteins 156
6 5 Epidermal, Hair and Nail Proteins 156
7 Biochemical Applications 157
7 1 Enzymology 157
711 Oxidoreductases 157
712 Transferases • • • • 158
713 Hydrolases 158
714 Lyases, Isomerases and Ligases -^ _ _ 160
7 2 Endocrinology 161
721 Pituitary Hormones 161
722 Insulin 162
7 3 Immunology 162
7 4 Animal Body Fluids 163
741 Animal Milk Proteins 163
742 Animal Serum Proteins 163
743 Other Animal Body Fluids 164
xvi Table of Contents
7 5 Neurology 165
7 6 Histology and Cytology (Animal Tissue and Cell Proteins) 165
761 Crystallin and other Ocular Lens Proteins 165
762 Collagen, Muscle and Cutaneous Proteins 166
763 Histones and other Nuclear Proteins 166
764 Other Subcellular Proteins 168
765 Membrane Proteins 168
7 7 Entomology 169
7 8 Botany 169
781 Seed Plants 169
782 Protozoa and Fungi 171
7 9 Microbiology 172
791 Bacteria - 172
792 Viruses and Phages • 176
7 10 Nucleic Acids 176
7 10 1 Special Conditions for Optimal Electrophoretic Sepa-
rations of RNA in Polyacrylamide Gel - 177
7 10 2 Low Molecular Weight RNA and Polynucleotides 178
7 10 3 High Molecular Weight RNA 180
7 10 4DNA and Polydeoxynucleotides 184
8 Applications in Food Technology and Forensic Analysis 185
References 187
List of Firms 211
List of Abbreviations 213
Subject Index 214
Fold-out Chart: Brief Instructions for the Preparation
of Gels and for the Electrophoretic Procedure
|
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| id | DE-604.BV000934527 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T15:21:29Z |
| institution | BVB |
| isbn | 3110034956 |
| language | English |
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| owner_facet | DE-19 DE-BY-UBM DE-M49 DE-BY-TUM DE-29 DE-29T DE-355 DE-BY-UBR DE-20 DE-83 DE-188 |
| physical | XVI, 222 S. Ill., graph. Darst. |
| publishDate | 1971 |
| publishDateSearch | 1971 |
| publishDateSort | 1971 |
| publisher | de Gruyter |
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| series2 | Working methods in modern science |
| spelling | Maurer, Hans Rainer Verfasser aut Diskelektrophorese Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables by H. Rainer Maurer 2., rev. and expanded ed. Berlin [u.a.] de Gruyter 1971 XVI, 222 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Working methods in modern science Electrophoresis cabt Disc-Elektrophorese Gelelektrophorese (DE-588)4123267-7 gnd rswk-swf Gelelektrophorese (DE-588)4123267-7 s DE-604 HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=000572044&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Maurer, Hans Rainer Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables Electrophoresis cabt Disc-Elektrophorese Gelelektrophorese (DE-588)4123267-7 gnd |
| subject_GND | (DE-588)4123267-7 |
| title | Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables |
| title_alt | Diskelektrophorese |
| title_auth | Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables |
| title_exact_search | Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables |
| title_full | Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables by H. Rainer Maurer |
| title_fullStr | Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables by H. Rainer Maurer |
| title_full_unstemmed | Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables by H. Rainer Maurer |
| title_short | Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis |
| title_sort | disc electrophoresis and related techniques of polyacrylamide gel electrophoresis with 16 tables |
| title_sub | with 16 tables |
| topic | Electrophoresis cabt Disc-Elektrophorese Gelelektrophorese (DE-588)4123267-7 gnd |
| topic_facet | Electrophoresis Disc-Elektrophorese Gelelektrophorese |
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