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Table of contents
- Cover
- Contentsvii
- Prefacev
- List of Contributorsxv
- Chapter 1. Overview of Proteome Analysis1
- 1. Introduction1
- 2. Scope of the Proteomics Problem2
- 3. Global versus Targeted Proteomics4
- 4. Top Down Protein Profiling Methods5
- 5. Bottom Up Protein Profiling Methods10
- 6. Automation, Miniaturization, and Future Prospects13
- 7. Summary14
- Acknowledgements15
- References15
- Chapter 2. Protein Profile Comparisons of Microorganisms, Cells and Tissues using 2D Gels19
- 1. Introduction20
- 2. Challenges of Protein Separation Methods for Proteome Analysis21
- 3. Current Technology of 2D Electrophoresis with IPGs (IPG-Dalt)38
- 4. 2D PAGE Databases64
- 5. Summary64
- Acknowledgements65
- References65
- Chapter 3. Protein Profiling using Two-Dimensional Gel Electrophoresis with Multiple Fluorescent Tag75
- 1. Introduction76
- 2. Mechanics of DIGE Technology77
- 3. Characterization of DIGE Technology80
- 4. MS Identification of Cyanine Labeled Proteins88
- 5. Summary89
- Acknowledgements90
- References90
- Chapter 4. Electrophoretic Prefractionation for Comprehensive Analysis of Proteomes93
- 1. Introduction94
- 2. Electrophoretic Prefractionation Methods96
- 3. Strategies for Analysis of Large Soluble Proteins and Insoluble Proteins103
- 4. Downstream Proteome Analysis After Sample Fractionation105
- 5. Summary114
- Acknowledgments115
- References115
- Chapter 5. Modification Specific Proteomics Applied to Protein Glycosylation and Nitration119
- 1. Introduction119
- 2. Glycosylation121
- 3. Tyrosine Nitration133
- 4. Summary134
- References134
- Chapter 6. Phosphoproteomics: Mass Spectrometry Based Techniques for Systematic Phosphoprotein Analy139
- 1. Introduction140
- 2. Modification-Specific Proteomics and Phosphoproteomics141
- 3. Detection and Sequencing of Phosphopeptides by Mass Spectrometry148
- 4. Cellular Dynamics: Integrated Methods for Quantitative Phosphoproteome Analysis154
- 5. Bioinformatics Tools for Phosphoprotein Sequence Analysis and Mass Spectrometry Data Interpretati155
- 6. Summary156
- Acknowledgements157
- References157
- Chapter 7. Protein Identification by In-Gel Digestion and Mass Spectrometry163
- 1. Introduction164
- 2. In-Gel Digestion165
- 3. Mass Spectrometry170
- 4. Troubleshooting176
- 5. Summary178
- Acknowledgements179
- References179
- Chapter 8. The Use of Accurate Mass and Time Tags Based Upon High-Throughput Fourier Transform Ion C183
- 1. Introduction184
- 2. Overall Experimental and Data Processing Approach186
- 3. Sample Processing188
- 4. High-Resolution Separations191
- 5. Chromatographic Separations Coupled to FTICR194
- 6. Generation of Accurate Mass and Time Tags and Their Utilization195
- 7. The Dynamic Range of Proteome Coverage196
- 8. Demonstration of Global Proteomic Characterization205
- 9. Overcoming Challenges to Proteome-wide Measurements211
- 10. Summary215
- Acknowledgements218
- References219
- Chapter 9. Clinical Applications of Proteomics225
- 1. Introduction225
- 2. Correlative Studies Using Proteomics and Transcriptomics226
- 3. Disease Marker Identification Using Proteomics228
- 4. Disease Tissue Analysis Using Proteomics231
- 5. Protein Microarrays as a Novel Technology for Disease Investigations234
- 6. Summary237
- References237
- Chapter 10. Affinity-based Biosensors, Microarrays and Proteomics243
- 1. Introduction244
- 2. Biosensor Technology246
- 3. Biosensor Applications253
- 4. Protein Chip Mass Spectrometry Using SELDI260
- 5. Protein Chips and Microarrays266
- 6. Summary275
- References275
- Chapter 11. Protein Expression Library Resources for Proteome Studies287
- 1. Introduction287
- 2. Clone Collections290
- 3. Summary300
- Acknowledgements300
- References301
- Chapter 12. Automation of Proteome Analysis305
- 1. Introduction306
- 2. Experimental Design307
- 3. Sample Preparation308
- 4. 2D PAGE311
- 5. Image Analysis312
- 6. Robotics for Cutting, Digestion and Spotting313
- 7. Protein Fingerprinting by MALDI MS314
- 8. Peptide Fingerprinting by MS/MS319
- 9. The Crucial Elements: LIMS and Data Mining321
- 10. Summary323
- Acknowledgements324
- References324
- Chapter 13. Micro- and Nanotechnology for Proteomics327
- 1. Introduction327
- 2. Benefits of Miniaturisation329
- 3. Miniaturisation in Proteomics330
- 4. Fabrication of Microstructures332
- 5. Microstructures for Proteomics335
- 6. Summary364
- Acknowledgements365
- References365
- Index371
Book details
- Vendor Elsevier S & T
- SKU 9780444510242
- ISBN-13 9780080515304
- Author Speicher, David W
- Category Science
- Subject Analytic
Do you have questions about this book?
This book explores the current status of proteomics, an exciting new discipline, which is less than 10 years old. This new field has rapidly grown into a major commercial and research enterprise with great prospects for dramatically advancing our knowledge of basic biological and disease processes. The contributors to this book are an international panel of proteomics experts, who review and discuss the current status of specific technologies and approaches.
Proteomics represents an exciting new way to pursue biological and biomedical science at an unprecedented pace. Proteomics takes a broad, comprehensive, systematic approach to understanding biology that is generally unbiased and not dependent upon existing knowledge. The major components of proteomics from basic discovery using a range of alternative analytical methods to discovery validation and use for clinical applications are discussed. State-of-the-art protein profiling methods include high resolution two-dimensional gels, two-dimensional differential in-gel electrophoresis, LC-MS and LC-MS/MS using accurate mass tags, and protein identifications of proteins from gels using mass spectrometry methods are discussed in depth. Other chapters describe comprehensive characterization of proteomes using electrophoretic prefractionation and analyses of sub-proteomes based on specific posttranslational modifications including the phospho-proteome, the glyco-proteome, and nitrated proteins. These conventional proteome analysis chapters are complemented by discussion of emerging technologies and approaches such as affinity based biosensor proteomics as well as the use of protein microarrays, microfluidics and nanotechnology. Strategies for improving throughput by automation are also discussed. Additional chapters address the application of current proteome techniques to clinical problems and the availability of protein expression library resources for proteome studies.
· Authored by international experts in the field
· Covers a wide range of topics including 2-D gels, global proteomics using accurate mass tags, global proteomics using electrophoretic prefractionation, microfluidics, and nanotechnology
· Includes state-of-the-art protein profiling methods, and emerging technologies
Proteomics represents an exciting new way to pursue biological and biomedical science at an unprecedented pace. Proteomics takes a broad, comprehensive, systematic approach to understanding biology that is generally unbiased and not dependent upon existing knowledge. The major components of proteomics from basic discovery using a range of alternative analytical methods to discovery validation and use for clinical applications are discussed. State-of-the-art protein profiling methods include high resolution two-dimensional gels, two-dimensional differential in-gel electrophoresis, LC-MS and LC-MS/MS using accurate mass tags, and protein identifications of proteins from gels using mass spectrometry methods are discussed in depth. Other chapters describe comprehensive characterization of proteomes using electrophoretic prefractionation and analyses of sub-proteomes based on specific posttranslational modifications including the phospho-proteome, the glyco-proteome, and nitrated proteins. These conventional proteome analysis chapters are complemented by discussion of emerging technologies and approaches such as affinity based biosensor proteomics as well as the use of protein microarrays, microfluidics and nanotechnology. Strategies for improving throughput by automation are also discussed. Additional chapters address the application of current proteome techniques to clinical problems and the availability of protein expression library resources for proteome studies.
· Authored by international experts in the field
· Covers a wide range of topics including 2-D gels, global proteomics using accurate mass tags, global proteomics using electrophoretic prefractionation, microfluidics, and nanotechnology
· Includes state-of-the-art protein profiling methods, and emerging technologies
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