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With the sequencing of the human genome complete, the field of proteomics, still in its early stages, has become an important and informative field of biomedical research. Many of the protocols and techniques commonly employed by proteomics researchers have been refined over the past several years, while at the same time, new and innovative methods have also been developed. Significantly, many of the protocols used in the study of the human proteome have been developed to examine proteins derived specifically from cell lines, body fluids or human tissues. As a consequence, not all protocols are compatible with the full-range of protein sources commonly used in the laboratory. The current volume on Tissue Proteomics seeks to bring together a number of useful and innovative protocols developed particularly for the proteomic profiling of human tissues. The first two protocols presented in this work focus on high-throughput g- based techniques. Chapter 1 describes 2-D DIGE, a method used to examine the entire proteome within discreet pI fractions, whereas Chapter 2 details several complementary techniques for the specific analysis of glycoproteins found in tissue samples. The next chapters, Chapters 3 and 4, focus on SELDI-MS and MALDI-TOF, two techniques that make use of mass spectrometry for the characterization and identification of proteins. These techniques may be used alone or in combination with other proteomic methods, such gel-based assays, where mass spectrometry is useful for additional characterization and identification of proteins.
List of contents
Preface.- Contributors.- 1 Capabilities Using 2-D DIGE in Proteomics Research: The New Gold Standard for 2-D Gel ElectrophoresisChristine R. Rozanas and Stacey M. Loyland.- 2 Protein Carbohydrate Analysis: Gel-Based Staining, Liquid Chromatography, Mass Spectrometry, and Microarray ScreeningInjae Shin, Alina D. Zamfir, and Bin Ye.- 3 Surface-Enhanced Laser Desorption/Ionization Mass Spectrometry for Protein and Peptide Profiling of Body FluidsAudrey Gagnon, Qian Shi, and Bin Ye.- 4 A Novel Approach Using MALDI-TOF/TOF Mass Spectrometry and Prestructured Sample Supports (AnchorChip Technology) for Proteomic Profiling and Protein IdentificationSau-Mei Leung and Rebecca L. Pitts.- 5 Automated Laser Capture Microdissection for Tissue ProteomicsAdrianna S. Rodriguez, Benjamin H. Espina, Virginia Espina, and Lance A. Liotta.- 6 Tissue Microarrays: An OverviewRajiv Dhir.- 7 Frozen Protein ArraysStephen M. Hewitt and Robert A. Star.- 8 Reverse Phase Protein Microarrays for Theranostics and Patient-Tailored TherapyVirginia Espina, Julia Wulfkuhle, Valerie S. Calvert, Lance A. Liotta, and Emanuel F. Petricoin III.- 9 Antibody Arrays for Determination of Relative Protein AbundancesGrigoriy S. Chaga.- 10 SH2 Domain-Based Tyrosine Phosphorylation ArrayXin Jiang, Lesile Roth, Stephanie Han, and Xianqiang Li.- 11 Immunoregulomics: A Serum Autoantibody-Based Platform for Transcription Factor ProfilingOliver W. Tassinari, Margarita Aponte, Robert J. Caiazzo Jr., and Brian C.-S. Liu.- 12 The "Reverse Capture" Autoantibody Microarray: An Innovative Approach to Profiling the Autoantibody Response to Tissue-Derived Native AntigensJoshua R. Ehrlich, Liangdan Tang, Robert J. Caiazzo Jr., Daniel W. Cramer, Shu-Kay Ng, Shu-Wing Ng, and Brian C.-S. Liu.- 13 Biorepository Standards andProtocols for Collecting, Processing, and Storing Human TissuesDean Troyer.
Summary
With the sequencing of the human genome complete, the field of proteomics, still in its early stages, has become an important and informative field of biomedical research. Many of the protocols and techniques commonly employed by proteomics researchers have been refined over the past several years, while at the same time, new and innovative methods have also been developed. Significantly, many of the protocols used in the study of the human proteome have been developed to examine proteins derived specifically from cell lines, body fluids or human tissues. As a consequence, not all protocols are compatible with the full-range of protein sources commonly used in the laboratory. The current volume on Tissue Proteomics seeks to bring together a number of useful and innovative protocols developed particularly for the proteomic profiling of human tissues. The first two protocols presented in this work focus on high-throughput g- based techniques. Chapter 1 describes 2-D DIGE, a method used to examine the entire proteome within discreet pI fractions, whereas Chapter 2 details several complementary techniques for the specific analysis of glycoproteins found in tissue samples. The next chapters, Chapters 3 and 4, focus on SELDI-MS and MALDI-TOF, two techniques that make use of mass spectrometry for the characterization and identification of proteins. These techniques may be used alone or in combination with other proteomic methods, such gel-based assays, where mass spectrometry is useful for additional characterization and identification of proteins.
