UID:
almafu_9958131492302883
Umfang:
1 online resource (915 p.)
Ausgabe:
2nd ed.
ISBN:
9786612754975
,
9781282754973
,
1282754971
,
9780080923178
,
0080923178
Serie:
Methods in enzymology ; v. 463
Inhalt:
The 2e of this classic Guide to Protein Purification provides a complete update to existing methods in the field, reflecting the enormous advances made in the last two decades. In particular, proteomics, mass spectrometry, and DNA technology have revolutionized the field since the first edition's publication but through all of the advancements, the purification of proteins is still an indispensable first step in understanding their function. This volume examines the most reliable, robust methods for researchers in biochemistry, molecular and cell biology, genetics, pharmacology and b
Anmerkung:
Previous ed.: 1990.
,
Front Cover; Methods in Enzymology: Guide to Protein Purification, 2nd Edition; Copyright Page; Contents; Contributors; Preface; Methods in Enzymology; Preface to Chapter 1; Chapter 1: Why Purify Enzymes?; Acknowledgment; Section 1: Developing Purification Procedures; Chapter 2: Strategies and Considerations for Protein Purifications; 1. General Considerations; 2. Source of the Protein; 3. Preparing Extracts; 4. Bulk or Batch Procedures for Purification; 5. Refined Procedures for Purification; 6. Conclusions; References; Chapter 3: Use of Bioinformatics in Planning a Protein Purification
,
1. What You Can Learn from an Amino Acid Sequence2. What You Cannot yet Predict; 3. Conclusion; References; Chapter 4: Preparing a Purification Summary Table; 1. Introduction; 2. The Importance of Footnotes; 3. The Value of an SDS-Polyacrylamide Gel Analysis on Main Protein Fractions; 4. Some Common Mistakes and Problems; Section 2: General Methods for Handling Proteins and Enzymes; Chapter 5: Setting Up a Laboratory; 1. Supporting Materials; 2. Detection and Assay Requirements; 3. Fractionation Requirements; Chapter 6: Buffers: Principles and Practice; 1. Introduction; 2. Theory
,
3. Buffer Selection4. Buffer Preparation; 5. Volatile Buffers; 6. Broad-Range Buffers; 7. Recipes for Buffer Stock Solutions; References; Chapter 7: Measurement of Enzyme Activity; 1. Introduction; 2. Principles of Catalytic Activity; 3. Measurement of Enzyme Activity; 4. Formulation of Reaction Assay Mixtures; 5. Discussion; Acknowledgments; References; Chapter 8: Quantitation of Protein; 1. Introduction; 2. General Instructions for Reagent Preparation; 3. Ultraviolet Absorption Spectroscopy; 4. Dye-Based Protein Assays; 5. Coomassie Blue (Bradford) Protein Assay (Range: 1-50 mug)
,
6. Lowry (Alkaline Copper Reduction Assays) (Range: 5-100 mug)7. Bicinchoninic Acid (BCA) (Range: 0.2-50 mug); 8. Amine Derivatization (Range: 0.05-25 mug); 9. Detergent-Based Fluorescent Detection (Range: 0.02-2 mug); 10. General Instructions; Acknowledgment; References; Chapter 9: Concentration of Proteins and Removal of Solutes; 1. Chromatography; 2. Electrophoresis; 3. Dialysis; 4. Ultrafiltration; 5. Lyophilization; 6. Precipitation; 7. Crystallization; References; Chapter 10: Maintaining Protein Stability; 1. Causes of Protein Inactivation; 2. General Handling Procedures
,
3. Concentration and Solvent Conditions4. Stability Trials and Storage Conditions; 5. Proteolysis and Protease Inhibitors; 6. Loss of Activity; Section 3: Recombinant Protein Expression and Purification; Chapter 11: Selecting an Appropriate Method for Expressing a Recombinant Protein; 1. Introduction; 2. Escherichia coli; 3. Pichia pastoris; 4. Baculovirus/Insect Cells; 5. Mammalian Cells; 6. Protein Characteristics; 7. Recombinant Protein Applications; 8. Conclusion; References; Chapter 12: Bacterial Systems for Production of Heterologous Proteins; 1. Introduction
,
2. Heterologous Protein Production Using Escherichia coli
,
English
Weitere Ausg.:
ISBN 9780123745361
Weitere Ausg.:
ISBN 0123745365
Sprache:
Englisch
Bookmarklink
