Title:
2D PAGE : sample preparation and fractionation
Series:
Methods in molecular biology
Publication Information:
Totowa, NJ : Humana Press, 2008
Physical Description:
v.
ISBN:
9781588297228
General Note:
Library has vol. 1
Added Author:
Available:*
Library | Item Barcode | Call Number | Material Type | Item Category 1 | Status |
|---|---|---|---|---|---|
Searching... | 30000010166664 | QH506 T96 2008 | Open Access Book | Book | Searching... |
On Order
Summary
Summary
This book, split into two volumes, presents a broad coverage of the principles and recent developments of sample preparation and fractionation tools in Expression Proteomics in general and for two-dimensional electrophoresis (2-DE) in particular. 2-DE, with its unique capacity to resolve thousands of proteins in a single run, is still a fundamental research tool for nearly all protein-related scientific projects. The methods described here in detail are not limited to 2-DE and can also be applied to other protein separation techniques. Because each biological sample is unique, a suited sample preparation strategy has to consider the type of sample as well as the type of biological question being addressed. The complex nature of proteins often requires a multitude of sample preparation options. In addition, sample preparation is not only a prerequisite for a successful and reproducible Proteomics exp- iment, but also the key factor to meaningful data evaluation. Interestingly, not much attention was paid to this area during Proteomics methodology dev- opment and therefore this book is intended to explain in depth how proteins from various sources can be properly isolated and prepared for reproducible Proteome analysis.
Table of Contents
| Preface | p. v |
| Contributors | p. xi |
| Part I Sample Preparation Basics | |
| 1 Mechanical/Physical Methods of Cell Disruption and Tissue Homogenization | p. 3 |
| 2 Bacteria and Yeast Cell Disruption Using Lytic Enzymes | p. 23 |
| 3 Sample Solublization Buffers for Two-Dimensional Electrophoresis | p. 35 |
| 4 Quantitation of Protein in Samples Prepared for 2-D Electrophoresis | p. 43 |
| 5 Removal of Interfering Substances in Samples Prepared for Two-Dimensional (2-D) Electrophoresis | p. 51 |
| 6 Protein Concentration by Hydrophilic Interaction Chromatography Combined with Solid Phase Extraction | p. 63 |
| Part II Protein Labeling Techniques | |
| 7 Difference Gel Electrophoresis Based on Lys/Cys Tagging | p. 73 |
| 8 Isotope-Coded Two-Dimensional Maps: Tagging with Deuterated Acrylamide and 2-Vinylpyridine | p. 87 |
| 9 Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) | p. 101 |
| 10 ICPL-Isotope-Coded Protein Label | p. 113 |
| 11 Radiolabeling for Two-Dimensional Gel Analysis | p. 125 |
| Part III Fractionation of Proteins by Chemical Reagents and Chromatography | |
| 12 Sequential Extraction of Proteins by Chemical Reagents | p. 139 |
| 13 Reducing Sample Complexity by RP-HPLC: Beyond the Tip of the Protein Expression Iceberg | p. 147 |
| 14 Enriching Basic and Acidic Rat Brain Proteins with Ion Exchange Mini Spin Columns Before Two-Dimensional Gel Electrophoresis | p. 157 |
| 15 Reducing the Complexity of the Escherichia coli Proteome by Chromatography on Reactive Dye Columns | p. 167 |
| 16 Reducing Sample Complexity in Proteomics by Chromatofocusing with Simple Buffer Mixtures | p. 187 |
| 17 Fractionation of Proteins by Immobilized Metal Affinity Chromatography | p. 205 |
| 18 Fractionation of Proteins by Heparin Chromatography | p. 213 |
| Part IV Fractionation of Proteins by Electrophoresis Methods | |
| 19 Fractionation of Complex Protein Mixtures by Liquid-Phase Isoelectric Focusing | p. 225 |
| 20 Microscale Isoelectric Focusing in Solution | p. 241 |
| 21 Prefractionation, Enrichment, Desalting and Depleting of Low Volume and Low Abundance Proteins and Peptides Using the MF10 | p. 257 |
| 22 Sample Prefractionation in Granulated Sephadex IEF Gels | p. 277 |
| 23 Free-Flow Electrophoresis System for Plasma Proteomic Applications | p. 287 |
| 24 Protein Fractionation by Preparative Electrophoresis | p. 301 |
| Part V Enrichment Strategies for Organelles, Multiprotein Complexes, and Specific Protein Classes | |
| 25 Isolation of Endocitic Organelles by Density Gradient Centrifugation | p. 317 |
| 26 Isolation of Highly Pure Rat Liver Mitochondria with the Aid of Zone-Electrophoresis in a Free Flow Device (ZE-FFE) | p. 333 |
| 27 Isolation of Proteins and Protein Complexes by Immunoprecipitation | p. 349 |
| 28 Isolation of Phosphoproteins | p. 365 |
| 29 Glycoprotein Enrichment Through Lectin Affinity Techniques | p. 373 |
| 30 Isolation of Bacterial Cell Membranes Proteins Using Carbonate Extraction | p. 397 |
| 31 Enrichment of Membrane Proteins by Partitioning in Detergent/Polymer Aqueous Two-Phase Systems | p. 403 |
| 32 The Isolation of Detergent-Resistant Lipid Rafts for Two-Dimensional Electrophoresis | p. 413 |
| 33 Isolation of Membrane Protein Complexes by Blue Native Electrophoresis | p. 423 |
| 34 Tissue Microdissection | p. 433 |
| Index | p. 449 |
