An efficient method for high-pH peptide fractionation based on C18 StageTips for in-depth proteome profiling

Literature Information

Publication Date 2019-08-20
DOI 10.1039/C9AY01269A
Impact Factor 2.896
Authors

Kisoon Dan, Hyunsuk Shin, Junghun Lee, Joseph Injae Wang, Dohyun Han


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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.

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