Protein phosphorylation controls many basic cellular processes, such as cell growth, differentiation, migration, metabolism, and cell death, and its study can provide key insights into the signal transduction pathways that are activated in cells in response to different stimuli, such as growth factor stimulation or exposure to toxicants. In Phospho-Proteomics: Methods and Protocols, expert researchers contribute both well-established protocols and some of the newest strategies for the identification and evaluation of protein phosphorylation on Tyr, Ser, and Thr residues, including topics such as 2-dimensional gel electrophoresis and protein phosphorylation, enrichment of phospho-proteins and peptides, quantitative analysis of phosphorylation by labeling and MS analysis, and antibody and kinase arrays. Comprehensive phospho-proteomics laboratory manual and research guide for both novice and experienced researchers Providesstep-by-step protocols written by leading researchers in the field that are fully adaptable to any specific phospho-proteome analysis task Includes two detailed critical reviews on qualitative and quantitative mass-spectrometry-basedphospho-proteomic analysis INDICE: From the contents A High-Resolution Two Dimensional Gel- and Pro-QDPS-Based Proteomics Workflow for Phosphoprotein Identification and Quantitative Profiling.- Differential Phospho-Protein Labeling (DIPPL) using 32P and 33P.- Identification of Oxidative Stress-Induced Tyrosine Phosphorylated Proteins by Immunoprecipitation and Mass Spectrometry.- Enrichment and Characterization of Phosphopeptides by Immobilized Metal Affinity Chromatography (IMAC) and Mass Spectrometry.- The Use of Titanium Dioxide Micro-Columns to Selectively Isolate Phosphopeptides from Proteolytic Digests.- Enrichment and Separation ofMono- and Multi-Phosphorylated Peptides Using Sequential Elution from IMAC prior to Mass Spectrometric Analysis.- Use of Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) for Phosphotyrosine Protein Identification and Quantitation.- Hydrophilic Interaction Chromatography for Fractionation and Enrichment of the Phosphoproteome.- SILAC for Global Phosphoproteomic Analysis.
- ISBN: 978-1-60327-833-1
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 265
- Fecha Publicación: 01/02/2009
- Nº Volúmenes: 1
- Idioma: Inglés