| Preface | p. ix |
| Preface to the Fourth Edition | p. xi |
| Acknowledgments | p. xiii |
| Abbreviations | p. xv |
| Daily Bread | p. 1 |
| Making Buffers | p. 1 |
| Protein Determination | p. 1 |
| BCA Assay | p. 2 |
| Bradford Assay | p. 3 |
| Lowry Assay | p. 3 |
| Starcher Assay | p. 3 |
| Protein Concentration | p. 4 |
| Gels | p. 4 |
| SDS Gels | p. 5 |
| For the Impatient: SDS Electrophoresis Without Stacker | p. 7 |
| Native Gels | p. 8 |
| Staining Gels | p. 9 |
| Fixing | p. 9 |
| Staining | p. 9 |
| Drying | p. 12 |
| Precipitation and Concentration | p. 13 |
| Denaturing Precipitation | p. 13 |
| Native Precipitation | p. 14 |
| Concentration | p. 14 |
| Blotting | p. 15 |
| Protein Staining on Blots | p. 17 |
| Blocking | p. 18 |
| Immunostaining | p. 19 |
| Ca[superscript 2+] Binding | p. 20 |
| Ligand Staining | p. 21 |
| Autoradiography of Gels and Blots | p. 22 |
| Ligand Binding | p. 25 |
| Radioactive Ligand Marking | p. 25 |
| Iodination of Peptides and Proteins | p. 26 |
| The Day After | p. 28 |
| Iodination of Molecules with Low MW | p. 29 |
| Isolation of Iodized Species | p. 29 |
| Advantages and Disadvantages of Iodination | p. 30 |
| Tritiation | p. 32 |
| Binding | p. 32 |
| Isolation of Membranes | p. 32 |
| Binding Assay | p. 34 |
| Binding Assays with Membranes | p. 37 |
| Development of Membrane Binding Assays | p. 38 |
| Binding Assays with Soluble Proteins | p. 39 |
| No Binding | p. 55 |
| Analysis of Binding Data | p. 57 |
| The Binding Reaction in Equilibrium | p. 57 |
| Kinetics | p. 69 |
| Cross-linking of Ligands | p. 71 |
| Three-component Cross-linking (3C Cross-linking) | p. 72 |
| Photoaffinity Cross-linking | p. 77 |
| Controls for Cross-linking Experiments | p. 79 |
| Purposes | p. 79 |
| Solubilization of Membrane Proteins | p. 83 |
| Detergents | p. 83 |
| Clean Concepts | p. 83 |
| Handling Detergents | p. 85 |
| Solubilization | p. 87 |
| Solubilization Criteria | p. 92 |
| Physical Parameters of Solubilized Membrane Proteins | p. 93 |
| Protein Detection via Functional Measurements | p. 95 |
| Translocators | p. 95 |
| Liposomes | p. 96 |
| Proteoliposomes | p. 97 |
| Reconstitution | p. 98 |
| Reconstitution from a Solution | p. 98 |
| Reconstitution in Preformed Liposomes | p. 100 |
| Flux Assay | p. 100 |
| Influx Assay | p. 102 |
| Efflux Assay | p. 104 |
| Constructive Thoughts | p. 105 |
| Cleaning and Purifying | p. 109 |
| Pure Fun | p. 109 |
| Conventional Purification Methods | p. 113 |
| The Column Technique | p. 113 |
| Purification Based on Size Differences | p. 114 |
| Purification Based on Charge Differences | p. 117 |
| Hydrophobic Chromatography | p. 124 |
| The Blue Gel | p. 125 |
| Affinity Chromatography | p. 125 |
| Lectin Chromatography | p. 125 |
| Ligand Chromatography | p. 127 |
| The Purity Test | p. 134 |
| Profiting | p. 134 |
| Antibodies | p. 137 |
| Production of Polyclonal Antibodies | p. 140 |
| Antigen | p. 140 |
| Adjuvant | p. 141 |
| Injection and Serum Harvesting | p. 142 |
| Purification of Antibodies | p. 143 |
| Immunoprecipitation | p. 145 |
| Immunoprecipitation with Immobilized Protein A | p. 145 |
| Immunoprecipitation with Immobilized Antibody | p. 146 |
| Immunoaffinity Chromatography | p. 148 |
| Antibodies Against Unpurified Proteins | p. 148 |
| Immunological Detection Techniques | p. 150 |
| Proteomics | p. 155 |
| Introduction | p. 155 |
| Sample Taking | p. 157 |
| 2D Gel Electrophoresis | p. 161 |
| Mass Spectroscopy of Peptides and Proteins | p. 165 |
| Mass Spectrometers | p. 165 |
| Sample Preparation for MALDI | p. 170 |
| The Possibilities of MALDI and ESI | p. 172 |
| Protein Chips | p. 174 |
| Protein Chips with SELDI | p. 174 |
| Fortune Cookies | p. 176 |
| Aptamers | p. 177 |
| Microsequencing | p. 179 |
| Preparing the Protein | p. 179 |
| Blocked N-Termini | p. 180 |
| Cleaving the Protein into Peptides | p. 181 |
| The Edman Degradation | p. 183 |
| Carboxyterminal Sequencing | p. 183 |
| Ladder Sequencing of Peptides | p. 184 |
| Strategy | p. 189 |
| Subunits | p. 191 |
| Number and Stoichiometry of Subunits | p. 191 |
| About the Difficulties with Stoichiometry Determinations | p. 191 |
| N&S with X-ray Structural Analysis | p. 192 |
| N&S with Hybridization Experiments | p. 192 |
| N&S with Cross-linking Experiments | p. 194 |
| N&S with Amino Acid Analyses or Antibodies | p. 199 |
| What Holds Our World Together? | p. 200 |
| Glycoproteins | p. 205 |
| How, Where, and for What Purpose Are Proteins Glycosylated? | p. 205 |
| Detecting Glycoproteins in Gels | p. 206 |
| Detection of Glycoproteins on Blots | p. 206 |
| Nonselective Glycoprotein Stain | p. 206 |
| Selective (Lectin) Stain | p. 208 |
| Deglycosylation | p. 208 |
| Glycosylation Inhibitor | p. 208 |
| Endoglycosidases | p. 210 |
| Chemical Deglycosylation | p. 214 |
| The Sugar Chains | p. 216 |
| Monosaccharide Composition | p. 216 |
| Structure and Sequence | p. 216 |
| Treasure Island: Writing Papers | p. 225 |
| Of the Paper | p. 225 |
| Of Writing a Paper | p. 225 |
| Desert Planet: Researching the Literature | p. 227 |
| Last Things | p. 228 |
| Professional Resources | p. 229 |
| Suppliers | p. 229 |
| Suppliers by Product | p. 229 |
| Index | p. 231 |
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