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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 2 illuminates specific applications of sample preparation tools and fractionation strategies to study a variety of biological systems.
Invaluable for students of Biochemistry, Biomedicine, Biology and Genomics and essential for the practicing scientists, 2D PAGE: Sample Preparation and Fractionation, Volumes 1 & 2 delves into the study of proteins with comprehensive and in depth methods and protocols to provide a greater understanding of this intricate and complicated field.
List of contents
Application of Fluorescence Dye Saturation Labeling for Differential Proteome Analysis of 1,000 Microdissected Cells from Pancreatic Ductal Adenocarcinoma Precursor Lesions.- Albumin and Immunoglobulin Depletion of Human Plasma.- Multi-Component Immunoaffinity Subtraction and Reversed-Phase Chromatography of Human Serum.- Immunoaffinity Fractionation of Plasma Proteins by Chicken IgY Antibodies.- Proteomics of Cerebrospinal Fluid: Methods for Sample Processing.- Sample Preparation of Bronchoalveolar Lavage Fluid.- Preparation of Nasal Secretions for Proteome Analysis.- Preparation of Urine Samples for Proteomic Analysis.- Isolation of Cytoplasmatic Proteins from Cultured Cells for Two-Dimensional Gel Electrophoresis.- Sample Preparation of Culture Medium from Madin-Darby Canine Kidney Cells.- Sample Preparation for Mass Spectrometry Analysis of Formalin-Fixed Paraffin-Embedded Tissue.- Metalloproteomics in the Molecular Study of Cell Physiology and Disease.- Protein Extraction from Green Plant Tissue.- The Terminator: A Device for High-Throughput Extraction of Plant Material.- Isolation of Mitochondria from Plant Cell Culture.- Isolation and Preparation of Chloroplasts from Arabidopsis thaliana Plants.- Isolation of Plant Cell Wall Proteins.- Isolation and Fractionation of the Endoplasmic Reticulum from Castor Bean (Ricinus communis) Endosperm for Proteomic Analyses.- Cell Wall Fractionation for Yeast and Fungal Proteomics.- Collection of Proteins Secreted from Yeast Protoplasts in Active Cell Wall Regeneration.- Sample Preparation Procedure for Cellular Fungi.- Isolation and Enrichment of Secreted Proteins from Filamentous Fungi.- Isolation and Solubilization of Cellular Membrane Proteins from Bacteria.- Isolation and Solubilization of Gram-Positive Bacterial Cell Wall-Associated Proteins.- Cell Fractionation of Parasitic Protozoa.
Summary
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. Step-by-step instructions alongside detailed illustrations enable you to quickly learn how to perform the methods yourself. Tricks of the trade and notes on troubleshooting and avoiding pitfalls help you 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.
