An efficient method for high-pH peptide fractionation based on C18 StageTips for in-depth proteome profiling. Issue 36 (30th August 2019)

• Record Type: Journal Article
• Authors: Kim, Hyeyoon; Dan, Kisoon; Shin, Hyunsuk; Lee, Junghun; Wang, Joseph Injae; Han, Dohyun
• Is Part Of: Analytical methods. Volume 11:Issue 36(2019)
• Page Start: 4693
• Page End: 4698
• Publication Date: 2019-08-30
• Subjects: Chemistry, Analytic -- Periodicals; Analytical biochemistry -- Periodicals; Chemical laboratories -- Standards -- Periodicals; 543.1905
• DOI: 10.1039/c9ay01269a
• Languages: English
• ISSNs: 1759-9660

Abstract

Abstract : Development of an efficient method for tip-based high-pH peptide fractionation suitable for proteomic analysis using small amounts of protein. Abstract : In-depth proteome profiling still relies widely on extensive off-line peptide fractionation to reduce the complexity of the proteome. However, the fractionation of protein samples in the sub-microgram range can be limited owing to sample loss during the fractionation process. Thus, it is essential to develop efficient microscale fractionation strategies. Here, we describe a modified method for high-pH peptide fractionation using C18-StageTips. Compared to a conventional strategy, our method generates improved peptide and protein identification rates under the same LC-MS/MS conditions. Finally, proteome profiling of sub-regions of the mouse brain suggested that our method can be used to analyze the proteome in-depth with limited amounts of protein samples.