4.8 Article

Simple Sodium Dodecyl Sulfate-Assisted Sample Preparation Method for LC-MS-Based Proteomics Applications

期刊

ANALYTICAL CHEMISTRY
卷 84, 期 6, 页码 2862-2867

出版社

AMER CHEMICAL SOC
DOI: 10.1021/ac203394r

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资金

  1. National Institutes of Health [R01 DK074795, RR018522]
  2. U.S. Department of Energy (DOE)
  3. DOE [DE-AC05-76RLO 1830]

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Sodium dodecyl sulfate (SDS) is one of the most popular laboratory reagents used for biological sample extraction; however, the presence of this reagent in samples challenges LC-MS-based proteomics analyses because it can interfere with reversed-phase LC separations and electrospray ionization. This study reports a simple SDS-assisted proteomics sample preparation method facilitated by a novel peptide-level SDS removal step. In an initial demonstration, SDS was effectively (>99.9%) removed from peptide samples through ion substitution-mediated DS- precipitation using potassium chloride (KCl), and excellent peptide recovery (>95%) was observed for <20 mu g of peptides. Further experiments demonstrated the compatibility of this protocol with LC-MS/MS analyses. The resulting proteome coverage obtained for both mammalian tissues and bacterial samples was comparable to or better than that obtained for the same sample types prepared using standard proteomics preparation methods and analyzed using LC-MS/MS. These results suggest the SDS-assisted protocol is a practical, simple, and broadly applicable proteomics sample processing method, which can be particularly useful when dealing with samples difficult to solubilize by other methods.

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