Targeted Phosphoproteomics Analysis of Immunoaffinity Enriched Tyrosine Phosphorylation in Mouse Tissue
نویسندگان
چکیده
Tyrosine kinases play a prominent role in transmitting and regulating various cell signaling pathways. Deregulation of this key PTM drives inappropriate proliferation and cell survival. MS-based global phosphoproteomics related studies have significantly evolved in recent years, but complexity of the signaling pathways and sub-stoichiometry levels of tyrosine phosphorylation sites, when compared to phosphoserine and phosphothreonine residues, limit extensive mapping of the phosphotyrosine (pTyr) repertoire1. Targeted phosphoprotein identification approaches offer a comprehensive solution to study selected sites involved in specific biochemical processes. More recently, coupling of peptide-level anti-pTyr immunoaffinity purification (IP) with LC/MS/MS has proven to be a good approach for profiling tyrosine phosphorylation2,3. This Application Note demonstrates a targeted pathway-level enrichment workflow from various mouse tissues, and shows the strength of the Agilent data analysis software including Spectrum Mill and GeneSpring for a pathway-centric based analysis of the phosphoproteome repertoire. Targeted Phosphoproteomics Analysis of Immunoaffinity Enriched Tyrosine Phosphorylation in Mouse Tissue
منابع مشابه
In-depth qualitative and quantitative profiling of tyrosine phosphorylation using a combination of phosphopeptide immunoaffinity purification and stable isotope dimethyl labeling.
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