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Full Description
This second edition provides up-to-date protocols focusing on techniques for the specific enrichment of phosphopeptides and phosphoproteins. Phosphoproteomics Methods and Protocols, Second Edition guides readers through different labeling strategies for quantitative phosphoproteomics; high-throughput mass spectrometry-based phosphoproteome analyses and phospho flow cytometry; and bioinformatics strategies for phosphoproteomics data analysis and integration. Additional protocols concentrate on the identification of kinase-substrate relationships by both high- and low-throughput approaches. 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, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Phosphoproteomics Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.
Contents
Thiol-ene Enabled Detection of Thiophosphorylation as a Labeling Strategy for Phosphoproteins.- Phosphopeptide Detection with Biotin-Labeled Phos-tag.- Phosphopeptide Enrichment by Covalent Chromatography after Solid Phase Derivatization of Protein Digests on Reversed Phase Supports.- Peptide Labeling Using Isobaric Tagging Reagents for Quantitative Phosphoproteomics.- Identification of Direct Kinase Substrates Using Analogue-Sensitive Alleles.- Quantitative Analysis of Tissue Samples by Combining iTRAQ Isobaric Labeling with Selected/Multiple Reaction Monitoring (SRM/MRM).- Enrichment Strategies in Phosphoproteomics.- Phosphopeptide Enrichment by Immobilized Metal Affinity Chromatography.- The use of Titanium Dioxide for Selective Enrichment of Phosphorylated Peptides.- Sequential Elution from IMAC (SIMAC) - An Efficient Method for Enrichment and Separation of Mono- and Multi-Phosphorylated Peptides.- Improving the Phosphoproteome Coverage for Limited Sample Amounts Using TiO2-SIMAC-HILIC (TiSH) Phosphopeptide Enrichment and Fractionation.- Offline High pH Reversed-Phase Peptide Fractionation for Deep Phosphoproteome Coverage.- Phosphopeptide Enrichment Using Various Magnetic Nanocomposites - An Overview.- Two Dimensional Gel Electrophoresis-Based Plant Phosphoproteomics.- Variable Digestion Strategies for Phosphoproteomics Analysis.- On-line LC-FAIMS-MS/MS for the Analysis of Phosphorylation in Proteins.- Simple and Reproducible Sample Preparation for Single Shot Phosphoproteomics With High Sensitivity.- Identification of Direct Kinase Substrates via Kinase Assay Linked Phosphoproteomics.- Phosphoprotein Detection by High-Throughput Flow Cytometry.- Resources for Assignment of Phosphorylation Sites on Peptides and Proteins.- From Phosphosites to Kinases.- Search Databases and Statistics - Pitfalls and Best Practices in Phosphoproteomics.- Systems Analysis for Interpretation of Phosphoproteomics Data.
