Tissue Proteomics

This book presents a compilation of methods that detail improved protein and peptide sample preparation and identification. Chapters guide readers through methods for depletion of myofibril-associated proteins, peptide sample preparation in urinary proteomics, purification of targeted proteins from native tissues, fractionation strategies for protein analysis, and GeLC-MS as a sample preparation method for sample preparation for proteomics using minimal amount of tissue. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Tissue Proteomics: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Depletion of Myofibril-Associated Proteins Using Selective Protein Extraction as a Tool in Cardiac Proteomics.- Discovery of Immune Reactive Human Proteins by High Density Protein Arrays and Customized Validation of Potential Biomarkers by ELISA.- HLA Class I and Class II-Induced Intracellular Signaling and Molecular Associations in Primary Human Endothelial Cells.- Characterization of Protein Complexes Using Chemical Cross-linking Coupled Electrospray Mass Spectrometry.- Optimization for Peptide Sample Preparation for Urine Peptidomics.- Purification of Target Proteins from Native Tissues: CCT Complex from Bovine Testes and PP2Ac from Porcine Brains.- Integrative Analysis of Proteomics Data to Obtain Clinically Relevant Markers.- Untargeted Screening of Urinary Peptides Using Off-line Nano-liquid Chromatography MALDI-TOF/TOF Mass Spectrometry.- Identifying Clinically Relevant Proteins for Targeted Analysis in the Development of a Multiplexed Proteomic Biomarker Assay.- In Situ Hybridization and Double Immunohistochemistry for the Detection of VEGF-A mRNA and CD34/Collagen IV Proteins in Renal Transplant Biopsies.- LC-SRM-based Targeted Quantification of Urinary Protein Biomarkers.- Fractionation of Soluble Proteins Using DEAE-, SP-, and Phenyl Sepharose Chromatographies for Proteomics.- GeLC-MS: A Sample Preparation Method for Proteomics Analysis of Minimal Amount of Tissue.- Targeted Proteomics Verification of Breast Cancer Biomarker Candidates.- Combination Strategy of Quantitative Proteomics Uncovers the Related-Proteins of Colorectal Cancer in the Interstitial Fluid of Colonic Tissue from the AOM-DSS Mouse Model.- Multiple Reaction Monitoring using Double Isotopologue Peptide Standards for Protein Quantification.- Isobaric Labeling Based LC-MS/MS Strategy for Comprehensive Profiling of Human Pancreatic Tissue Proteome.- MALDI Imaging Mass Spectrometry of N-glycans and Tryptic Peptides from the same Formalin-Fixed, Paraffin-embedded Tissue Section.- Differential Adipose Tissue Proteomics.- Quantification of Breast Cancer Protein Biomarkers at Different Expression Levels in Human Tumors.- Quantitative Proteomic Analysis of Mass Limited Tissue Samples for Spatially-resolved Tissue Profiling.- High-throughput Proteomics Analysis of Fresh-frozen Biopsy Tissue Samples using Pressure Cycling Technology Coupled with SWATH Mass Spectrometry.- Straightforward Protocol for Gel-free Proteomic Analysis of Adipose Tissue.- MALDI Imaging Combined to Laser Microdissection-based Microproteomics for Protein Identification. Application to Intratumor Heterogeneity Studies.