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2D PAGE: Sample Preparation and Fractionation

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Beschreibung

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.

Contains step-by-step instructions that ensure successful results

Provides tricks of the trade and notes on troubleshooting and avoiding known pitfalls



Autorentext
Focusing on two-dimensional electrophoresis (2-DE), this book sets forth the principles and recent developments of sample preparation and fractionation tools in expression proteomics. 2-DE, with its unique capacity to resolve thousands of proteins in a single run, remains a fundamental research tool for nearly all protein-related scientific projects. Step-by-step instructions alongside detailed illustrations enable biochemists, molecular and cellular biologists, and protein scientists to quickly learn how to perform the methods. Tricks of the trade and notes on troubleshooting and avoiding pitfalls help ensure successful results every time. The methods covered in this book are not limited to 2-DE; they can also be applied to a broad range of protein separation techniques.

Klappentext

The complex nature of proteins often requires a multitude of sample preparation options. In 2D PAGE: Sample Preparation and Fractionation, Volumes 1 & 2, Dr. Anton Posch presents a broad coverage of the principles and recent developments of sample preparation and fractionation tools in Expression Proteomics and specifically for two-dimensional electrophoresis (2-DE). Detailed laboratory methods describe the process of sample fractionation used to reduce the complexity of the protein sample and the methods of 2-DE analysis and a broad variety of gel-free mass spectrometry-based technologies to detect previously undetectable proteins. Vital information regarding protein isolation and preparation for reproducible proteome analysis previously overlooked by scientists has been clearly laid out in this pioneering two-volume set. Volume 1 focuses on the basics of sample preparation and protein labeling techniques with an emphasis on fractionation by chemical reagents, chromatography, and electrophoresis methods. The inclusion of meticulous enrichment strategies further aids the protein researcher.

 

Invaluable for students of Biochemistry, Biomedicine, Biology and Genomics and essential for the practicing scientists, 2D PAGE: Sample Preparation and Fractionation, Volumes 1 & 2 delve into the study of proteins with comprehensive and in depth methods and protocols to provide a greater understanding of this intricate and complicated field.



Inhalt
Sample Preparation and Fractionation: Volume 1 Contents Part I Sample preparation basics 1 MECHANICAL/PHYSICAL METHODS OF CELL DISRUPTION AND TISSUE HOMOGENIZATION (Stanley Goldberg, unusual chapter format!!!!) 2 Bacteria and yeast cells disruption using lytic enzymes (Oriana Salazar) 3 Sample solubilization buffers for two-dimensional electrophoresis (Walter Weiss, unusual chapter format!!!!) 4 Quantitation of protein in samples prepared for 2-D electrophoresis (Thomas Berkelman) 5 Removal of Interfering Substances in Samples Prepared for 2-D Electrophoresis (Thomas Berkelman) 6 Protein Concentration by Hydrophilic Interaction Chromatography combined with Solid Phase Extraction (Ulrich Schneider) Part II Protein labeling techniques 7 Difference Gel Electrophoresis Based on Lys/Cys Tagging (Reiner Westermeier) 8 Isotope-coded two-dimensional maps: tagging with deuterated acrylamide and 2- vinylpyridine (Pier Giorgio Righetti, unusual chapter format!!!!) 9 Stable Isotope Labeling by Amino acids in Cell culture (SILAC) (Albrecht Gruhler) 10 ICPL - Isotope-coded protein label (Josef Kellermann) 11 Radiolabeling for Two-Dimensional Gel Analysis (Hélian Boucherie) Part III Fractionation of proteins by chemical reagents and chromatography 12 Sequential extraction of proteins by chemical reagents (Stuart Cordwell) 13 Reducing sample complexity by RP-HPLC: beyond the tip of the protein expression iceberg (Gert van den Bergh) 14 Enriching Basic and Acidic Rat Brain Proteins with Ion Exchange Mini Spin Columns Prior To Two-Dimensional Gel Electrophoresis (Ning Liu) 15 Reducing the complexity of the Escherichia coli proteome by chromatography on reactive dye columns (Phil Cash) 16 Reducing sample complexity in Proteomics by Chromatofocusing with simple buffer mixtures (Douglas D.Frey) 17 Fractionation of Proteins by Immobilized Metal Affinity Chromatography (Qing-Yu He) 18 Fractionation of proteins by heparin chromatography (Qing-Yu He) Part IV Fractionation of proteins by electrophoresis methods 19 Fractionation of Complex Protein Mixtures By Liquid-Phase Isoelectric Focusing (Julie Hey) 20 Microscale isoelectric focusing in solution. A Method for Comprehensive and Quantitative Proteome Analysis Using 1-D and 2-D DIGE Combined with MicroSol IEF Prefractionation. (David Speicher) 21 Prefractionation, enrichment, desalting and depleting of low volume and low abundance proteins and peptides using the MF10 (Valerie Wasinger) 22 Sample Prefractionation in Granulated Sephadex IEF Gels (Angelika Görg) 23 Free-Flow Electrophoresis System for Plasma Proteomic Applications (Robert Wildgruber) 24 Protein fractionation by preparative electrophoresis (Michael Fountoulakis) Part V Enrichment strategies for organelles, multiprotein complexes and specific protein classes 25 Isolation of endocitic organelles by density gradient centrifugation (Lukas A. Huber) 26 Isolation of highly pure rat liver mitochondria with the aid of zone-electrophoresis in a free flow device (ZE-FFE) (Hans Zischka) 27 Isolation of Proteins and Protein Complexes by Immunoprecipitation (Barbara Kaboord) 28 Isolation of Phosphoproteins (Lynn A. Megeney) 29 Glycoprotein Enrichment through Lectin Affinity Techniques (Milos V. Novotny) 30 Isolation of Bacterial Cell Membranes Proteins using Carbonate Extraction (Mark Molloy) 31 Enrichment of Membrane Proteins by Partitioning in Detergent/Polymer Aqueous Two-Phase Systems (Folke Tjerneld) 32 The isolation of detergent-resistant lipid rafts for two-dimensional electrophoresis (Young-Gyu Ko) 33 Isolation of membrane protein complexes by Blue Native Electrophoresis (Lutz A.

Produktinformationen

Titel: 2D PAGE: Sample Preparation and Fractionation
Untertitel: Volume 1
Autor:
EAN: 9781588297228
ISBN: 1588297225
Format: Fester Einband
Herausgeber: Humana Press
Anzahl Seiten: 476
Gewicht: 875g
Größe: H241mm x B160mm x T30mm
Jahr: 2008
Untertitel: Englisch
Auflage: 2008

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