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Optimization of reversed-phase solid-phase extraction for shotgun proteomics analysis By Fanni Bugyi,Lilla Turiák,László Drahos and Gábor Tóth
Authors:Fanni Bugyi  Lilla Turiák  László Drahos  Gábor Tóth
Institution:1. MS Proteomics Research Group, Research Centre for Natural Sciences, Magyar tudósok körútja 2, Budapest, 1117 Hungary

Hevesy György PhD School of Chemistry, Eötvös Loránd University, Pázmány Péter sétány 1/a, Budapest, 1117 Hungary;2. MS Proteomics Research Group, Research Centre for Natural Sciences, Magyar tudósok körútja 2, Budapest, 1117 Hungary

Abstract:Reversed-phase solid-phase extraction (SPE) is the method of choice for the purification of proteomics samples. Even though the efficacy of SPE methods is sample type-dependent, the manufacturers' protocols are used in most studies. Using an optimized SPE method can lead to a substantial gain in identification and recovery. In this tutorial, we give a brief introduction to the most important parameters influencing SPE performance, and we present a short workflow (16 measurements) for optimizing the SPE procedure. This is complemented by method performance assessment instructions and a short troubleshooting guide to help users further understand and investigate their SPE methods.
Keywords:C18  desalting  HPLC  mass spectrometry  peptide  proteomics  purification  reversed phase  solid-phase extraction
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