Holtzhauer Basic Methods for the Biochemical Lab

1. Auflage 2006
ISBN: 978-3-540-32786-8
Verlag: Springer Berlin Heidelberg
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 252 Seiten

Reihe: Springer Lab Manuals

This book presents proven lab procedures and practical hints for research in analytical and preparative biochemistry, and offers convenient key data in numerous tables. Coverage includes quantitative methods; electrophoresis; chromatographic protocols; immunochemical protocols; centrifugation; and radioactivity. In additional chapters, tables offer quick access to a broad array of useful information, including SI units conversion factors; detergent, protein and nucleotide data; and the basic principles of statistics and enzyme and receptor kinetics are reviewed. This first English-language edition of a successful German-language manual is a valuable resource for students and working professionals in biochemistry, biotechnology and biomedical laboratories.

Autoren/Hrsg.

Holtzhauer, Martin

Inhaltsverzeichnis

1;Preface;6
2;Table of Contents;7
3;Abbreviations;14
4;1 Quantitative Methods;16
4.1;1.1 Quantitative Determinations of Proteins;16
4.2;1.2 Quantitative Determination of Nucleic Acids;28
4.3;1.3 Quantitative Phosphate Determinations;32
4.4;1.4 Monosaccharide Determination;34
4.5;1.5 Calculations in Quantitative Analysis;35
5;2 Electrophoresis;38
5.1;2.1 Polyacrylamide Gel Electrophoresis Systems;38
5.2;2.2 Agarose and Paper Electrophoresis;60
5.3;2.3 Aid in Electrophoresis;64
5.4;2.4 Staining Protocols;68
5.5;2.5 Electroelution from Gels;81
5.6;2.6 Drying of Electrophoresis Gels;94
5.7;2.7 Autoradiography of Radioactive Labeled Compounds in Gels;95
6;3 Chromatography;98
6.1;3.1 Thin-Layer Chromatography;98
6.2;3.2 Hints for Column Chromatography of Proteins;104
6.3;3.3 Gel Permeation Chromatography ( GPC; Gel Filtration, GF; Size- Exclusion Chromatography, SEC);108
6.4;3.4 Ion Exchange Chromatography (IEC);117
6.5;3.5 Hydrophobic Interaction Chromatography (HIC);122
6.6;3.6 Affinity Chromatography (AC);124
6.7;3.7 Concentration of Diluted Protein Solutions;139
7;4 Immunochemical Protocols;144
7.1;4.1 Conjugation of Haptens (Peptides) to Carrier Proteins;144
7.2;4.2 Immunization of Laboratory Animals;158
7.3;4.3 Ammonium Sulfate Fractionation of Immunoglobulins;159
7.4;4.4 Removal of Unspecific Immunoreactivities;161
7.5;4.5 Preparation of Egg Yolk IgY Fraction;163
7.6;4.6 Antibody Fragmentation;164
7.7;4.7 HEIDELBERGER Curve (Precipitin Curve);165
7.8;4.8 OUCHTERLONY Double-Radial Immunodiffusion;166
7.9;4.9 Immunoprecipitation of Antigens;168
7.10;4.10 Immunoelectrophoresis;169
7.11;4.11 Counterelectrophoresis;170
7.12;4.12 Dot-Blot Assay;171
7.13;4.13 Enzyme Immunosorbent Assay (EIA, ELISA);172
8;5 Centrifugation;176
8.1;5.1 Speed vs Centrifugal Force Graphs;176
8.2;5.2 Differential Centrifugation;179
8.3;5.3 Density Gradient Centrifugation;180
9;6 Radioactive Labeling;196
9.1;6.1 Radioactive Decay;197
9.2;6.2 Decay Tables for 32-Phosphorus, 35-Sulfur, and 125- Iodine;198
9.3;6.3 Enzymatic [32P]-Phosphate Incorporation into Proteins;200
9.4;6.4 Iodination with [125I]-Iodine Reagents;202
9.5;6.5 Scintillation Cocktails for Liquid Scintillation Counting;203
10;7 Buffers;206
10.1;7.1 Theoretical Considerations;206
10.2;7.2 Plot for Buffer Calculations;213
10.3;7.3 pH Indicators;214
10.4;7.4 Buffer Recipes;214
11;8 Tables;224
11.1;8.1 Concentration Units;224
11.2;8.2 Conversion Factors for SI Units;225
11.3;8.3 Data of Frequently Used Substances;227
11.4;8.4 Protein Data;231
11.5;8.5 Protease Inhibitors;236
11.6;8.6 Single-Letter Codes and Molecular Masses of Amino Acids;237
11.7;8.7 Spectroscopic Data of Nucleotides;240
11.8;8.8 Detergents (“Surfactants”);240
11.9;8.9 Refractive Index and Density of Sucrose Solutions;243
11.10;8.10 Ammonium Sulfate Saturation Table;244
11.11;8.11 Diluted Solutions;246
11.12;8.12 Mixture Rule;247
12;9 Statistics and Data Analysis;248
12.1;9.1 Statistical Equations;248
12.2;9.2 Data Analysis;252
12.3;9.3 Diagnostic Sensitivity and Specificity;259
12.4;9.4 Software for the Lab;259
13;Subject Index;262