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.
Provides time tested, reproducible laboratory methods and protocols authored by experts in the field, including tips and hints
Presents the most up to date and useful proteomic techniques for profiling human tissue
Provides indispensable protocols for biomarker discovery, therapeutic response profiling, and high throughput based techniques
With the completion of human genome sequencing, the field of proteomics has grown vital to biomedical research. Among the array of protocols developed to study the human proteome, many protocols examine proteins derived specifically from cell lines, body fluids or human tissues - not necessarily compatible with the full range of protein sources commonly used in the laboratory. Tissue Proteomics constitutes in one volume useful and innovative protocols developed specifically for the proteomic profiling of human tissues. A unique resource for researchers, Tissue Proteomics provides high-throughput gel-based techniques, microarrays and a number of other methods used in proteomic research, a field still in its early stages. This important book will prove indispensable to investigators of biomarker discovery and therapeutic response profiling, as well as those forging new paths in the fields of theranostics and personalized medicine.
Inhalt Preface 1. Capabilities using 2-D DIGE in Proteomics Research: The New Gold Standard for Two-dimensional Gel Electrophoresis. Christine R. Rozanas and Stacey M. Loyland 2. Protein Carbohydrate Analysis: Gel-based Staining, Liquid Chromatography, Mass Spectrometry and Microarray Screening. Injae Shin, Alina D. Zamfir, and Bin Ye 3. Surface-Enhanced Laser Desorption/Ionization Mass Spectrometry (SELDI-MS) for Protein and Peptide Profiling of Body Fluids. Audrey 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 Identification. Sau-Mei Leung, and Rebecca L. Pitts 5. Automated Laser Capture Microdissection for Tissue Proteomics. Adrianna S. Rodriguez, Benjamin H. Espina, Virginia Espina, and Lance A. Liotta. 6. Tissue Microarrays: An overview. Rajiv Dhir 7. Frozen Protein Arrays. Stephen M. Hewitt and Robert A. Star. 8. Reverse Phase Protein Microarrays for Theranostics and Patient Tailored Therapy. Virginia Espina, Julia Wulfkuhle, Valerie S. Calvert, Lance A. Liotta, and Emanuel F. Petricoin III 9. Antibody Arrays for Determination of Relative Protein Abundances. Grigoriy S. Chaga 10. SH2 Domain Based Tyrosine Phosphorylation Array. Xin Jiang, Leslie Rose, Stephanie Han, and Xianqiang Li 11. Immunoregulomics: a serum autoantibody-based platform for transcription factor profiling. Oliver 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 antigens. Joshua 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 and Protocols for Collecting, Processing, and Storing Human Tissues. Dean Troyer