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.
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.
