SILAC

This detailed volume explores the most recent methodologies and applications of stable isotope labeling by amino acids in cell culture (SILAC). With the development of new technologies in the field of mass spectrometry and bioinformatics, SILAC-based methods have become the first tool of choice in quantitative proteomics either on its own or in combination with other more recently developed approaches, which is elucidated in the chapters of this book. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, SILAC: Methods and Protocols serves as an ideal guide for researchers working in the area of functional proteomics and other aspects of protein science.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Contenu
Dynamic SILAC to Determine Protein Turnover in Neurons and Glia.- SILAC-Based Proteomic Analysis of Meiosis in the Fission Yeast Schizosaccharomyces pombe .- Identification of Protein Interaction Partners in Bacteria Using Affinity Purification and SILAC Quantitative Proteomics.- pSILAC-Based Determination of Cellular Protein Sorting into Extracellular Vesicles.- Quantification of Protein Palmitoylation by Cysteine-SILAC.- Comparative SUMO Proteome Analysis Using Stable Isotopic Labeling by Amino Acids (SILAC).- A Super-SILAC Approach for Profiling Histone Post-Translational Modifications.- SILAC-Based Quantitative Phosphoproteomics in Yeast.- Phosphotyrosine Profiling Using SILAC.- Chemoproteomic Profiling of Geranyl Pyrophosphate-Binding Proteins.- Quantitative Analysis of the Endoplasmic Reticulum-Associated Proteins Using ER-Localizable Reactive Molecules.- Workflow for Quantitative Proteomic Analysis of Intestinal Organoids Using SILAC.- Stable Isotope Labeling by Amino Acids and Bioorthogonal Noncanonical Amino Acid Tagging in Cultured Primary Neurons.- Heavy Methyl SILAC Metabolic Labeling of Human Cell Lines for High-Confidence Identification of R/K-Methylated Peptides by High-Resolution Mass Spectrometry.- SILAC-Based Quantitative Proteomic Analysis of Drosophila Embryos.- Super-SILAC Quantitative Proteome Profiling of Zebrafish Larvae.- Use of Nuclear and Chromatin Enrichment Procedures for Quantitation of Yeast DNA Replication Proteins Using SILAC.- Combination of Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and Substrate Trapping for the Detection of Transient Protein Interactions.- Profiling of Secreted Proteins in Serum-Containing Media Using BONCAT and Pulsed SILAC.- Using SILAC to Develop Quantitative Data Independent Acquisition (DIA) Proteomics Methods.- SILAC-IodoTMT for Assessment of the Cellular Proteome and Its Redox Status.- Targeted Absolute Protein Quantification Using SILAC Internal Standard and Full-LengthProtein Calibrators (TAQSI).