Protein Analysis and Purification Benchtop Techniques /
| Autor principal: | |
|---|---|
| Autor Corporativo: | |
| Formato: | eBook |
| Lenguaje: | English |
| Publicado: |
Boston, MA :
Birkhäuser Boston : Imprint: Birkhäuser,
1996.
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| Edición: | 2nd ed. 1996. |
| Materias: | |
| Acceso en línea: | https://doi.org/10.1007/978-1-4612-2056-5 |
Tabla de Contenidos:
- 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.