Protein Analysis and Purification Benchtop Techniques - Rosenberg, Ian M.; SpringerLink (Online service) (Boston, MA : Birkhäuser Boston : Imprint: Birkhäuser, 1996)
1 An Overview of This Manual -- Kits, Cores, and Computers -- How to Use This Book -- Basic Laboratory Equipment -- Laboratory Automation -- Beyond Protein Analysis and Purification -- 2 Protein Structure -- A. The Amino Acids -- B. The Four Levels of Protein Structure -- C. Chemical Characteristics of Proteins -- 3 Tracking the Target Protein -- A. Labeling Cells and Proteins -- B. Lysis-Preparation of the Cell Free Extract -- C. Principles of Immunoprecipitation -- D. Additional Methods to Identify Associated Proteins -- 4 Electrophoretic Techniques -- to Polyacrylamide Gel Electrophoresis (PAGE) -- A. Preparation of SDS-Polyacrylamide Gels -- B. 2-Dimensional (2-D) Gel Systems -- D. Recovery of Proteins from the Gel -- E. Identification of Enzyme Activity in Polyacrylamide Gels -- 5 Getting Started with Protein Purification -- A. Making a Cell Free Extract -- B. Protein Quantitation -- C. Manipulating Proteins in Solution -- D. Precipitation Techniques -- 6 Membrane Proteins -- A. Peripheral Membrane Proteins -- B. Integral Membrane Proteins -- C. Detergents -- 7 Transfer and Detection of Proteins on -- Membrane Supports -- A. Transfer of Proteins to Membrane Supports -- B. Staining the Blot -- C. Recovery of Proteins from the Blot -- D. Detection of the Target Protein -- 8 Identification of the Target Protein -- A. Peptide Mapping -- B. Enzymatic Cleavage of Proteins -- C. Chemical Cleavage -- D. Microsequencing from PVDF Membranes -- MS Basics -- Protein Identification by MS -- Protein Database Searches -- The Rise of Biological Databases -- 9 Identifying and Analyzing Posttranslational -- Modifications -- A. Glycosylation -- B. Phosphorylation -- C. Lipid Modification of Proteins -- D. Selected Modifications -- 10 Chromatography -- A. Important Terminology Used in Chromatography -- B. Gel Filtration Chromatography -- C. Introduction to HPLC -- D. Ion Exchange Chromatography: Separation on the Basis of Charge -- E. Hydrophobic Interaction Chromatography (HIC) -- Flow Rate -- 11 Recombinant Protein Techniques -- Recombinant Protein for Antibody Production -- Protein for Biochemical or Cell Biological Studies -- A. In vitro Transcription and Translation -- B. Recombinant Gene Products in E. coli: Expression, Identification and Characterization -- D. Expression of Foreign Proteins in Eukaryotic Cells -- Appendices -- A. Safety Considerations -- First Aid: Emergency Procedures -- B. Antibody Preparation -- Production of Polyclonal Antisera in Rabbits -- Purification of Antibody Using Protein A Affinity Columns -- Numbering Mice -- Molarity -- Choosing and Preparing Buffers -- Common Laboratory Solutions -- Extinction Coefficients -- D. Nucleic Acids -- Spectrophotometric Conversions -- DNA/Protein Conversions -- Oligonucleotide Concentrations -- RNA Precipitation -- E. Modifications and Motifs -- Nomenclature -- Protein Modification Sequences -- Protein Kinase Recognition Sequence Motifs -- Subcellular Localization Motifs -- Protein Databases -- F. Centrifugation -- Nomogram -- General Purpose Centrifuge Rotors -- Ultracentrifuge Rotors -- G. Proteases and Proteolytic Enzyme Inhibitors -- Commonly Used Proteases -- Protease Inhibitors -- H. Radioactivity -- Manual and Machine Film Processing -- L Tissue Culture -- Transwell Permeable Supports -- J. Miscellaneous -- Unit Prefixes -- The Greek Alphabet -- Abbreviations -- HPLC Pump Pressure Conversion -- Dipeptide Masses -- Mass Differences Considered in Molecular Weight Analysis of Proteins -- K. List of Suppliers, Vendors, Manufacturers.
Keywords: Proteins .; Biochemistry; Life sciences; Protein Science; Biochemistry, general; Life Sciences, general