2D PAGE: Sample Preparation and Fractionation

1. Fractionation of Proteins by Chemical Reagents and Chromatography

   1. Fractionation of Proteins by Immobilized Metal Affinity Chromatography

      • Xuesong Sun, Jen-Fu Chiu, Qing-Yu He

      Pages 205-212

   2. Fractionation of Proteins by Heparin Chromatography

      • Sheng Xiong, Ling Zhang, Qing-Yu He

      Pages 213-221

2. Fractionation of Proteins by Electrophoresis Methods

   1. Front Matter

      Pages 223-223

      PDF

   2. Fractionation of Complex Protein Mixtures by Liquid-Phase Isoelectric Focusing

      • Julie Hey, Anton Posch, Andrew Cohen, Ning Liu, Adrianna Harbers

      Pages 225-239

   3. Microscale Isoelectric Focusing in Solution

      • Mee-Jung Han, David W Speicher

      Pages 241-256

   4. Prefractionation, Enrichment, Desalting and Depleting of Low Volume and Low Abundance Proteins and Peptides Using the MF10

      • Valerie Wasinger, Linda Ly, Anna Fitzgerald, Brad Walsh

      Pages 257-275

   5. Sample Prefractionation in Granulated Sephadex IEF Gels

      • Angelika Görg, Carsten Lück, Walter Weiss

      Pages 277-286

   6. Free-Flow Electrophoresis System for Plasma Proteomic Applications

      • Robert Wildgruber, Jizu Yi, Mikkel Nissum, Christoph Eckerskorn, Gerhard Weber

      Pages 287-300

   7. Protein Fractionation by Preparative Electrophoresis

      • Michael Fountoulakis, Ploumisti Dimitraki

      Pages 301-313

3. Enrichment Strategies for Organelles, Multiprotein Complexes, and Specific Protein Classes

   1. Front Matter

      Pages 315-315

      PDF

   2. Isolation of Endocitic Organelles by Density Gradient Centrifugation

      • MarianaEÇaGuimarães Araùjo, Lukas Alfons Hube, Taras Stasyk

      Pages 317-331

   3. Isolation of Highly Pure Rat Liver Mitochondria with the Aid of Zone-Electrophoresis in a Free Flow Device (ZE-FFE)

      • Hans Zischka, Josef Lichtmannegger, Nora Jägemann, Luise Jennen, Daniela Hamöller, Evamaria Huber et al.

      Pages 333-348

   4. Isolation of Proteins and Protein Complexes by Immunoprecipitation

      • Barbara Kaboord, Maria Perr

      Pages 349-364

   5. Isolation of Phosphoproteins

      • Lawrence G. Puente, Lynn A. Megeney

      Pages 365-372

   6. Glycoprotein Enrichment Through Lectin Affinity Techniques

      • Yehia Mechref, Milan Madera, Milos V Novotny

      Pages 373-396

   7. Isolation of Bacterial Cell Membranes Proteins Using Carbonate Extraction

      • Mark P Molloy

      Pages 397-401

   8. Enrichment of Membrane Proteins by Partitioning in Detergent/Polymer Aqueous Two-Phase Systems

      • Henrik Everberg, Niklas Gustavasson, Folke Tjerned

      Pages 403-412

   9. The Isolation of Detergent-Resistant Lipid Rafts for Two-Dimensional Electrophoresis

      • Ki-Bum Kim, Jae-Seon Lee, Young-Gyu Ko

      Pages 413-422

   10. Isolation of Membrane Protein Complexes by Blue Native Electrophoresis

       • Veronika Reisinger, Lutz A. Eichacker

       Pages 423-431

   11. Tissue Microdissection

       • Heidi S. Erickson, John W. Gillespie, Michael R. Emmert-Buck

       Pages 433-448

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.

• Amino acid
• Escherichia coli
• Lipid
• Organelle
• Polyacrylamid-Gelelektrophorese
• Proteomics
• biochemistry
• biomedicine
• enzymes
• glycoprotein
• membrane proteins
• polymer
• protein
• protein complexes
• proteins

• Bio-Rad Laboratories GmbH, Germany

  Anton Posch

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