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  • Title: Characterization of zebrafish cardiac proteome using online pH gradient SCX-RP HPLC-MS/MS platform.
    Author: Zhang J, Lanham KA, Heideman W, Peterson RE, Li L.
    Journal: Methods Mol Biol; 2013; 1005():119-27. PubMed ID: 23606253.
    Abstract:
    Two-dimensional HPLC coupled with tandem MS (MS/MS) has become a mainstream technique in the shotgun proteomics for large-scale identification of proteins from biological samples. This powerful technology provides speed, sensitivity, and dynamic range which are essential to probe complex peptide mixtures from proteomic samples. Herein we present a pH gradient SCX-RP 2D HPLC-MS/MS method designed to improve the peptide resolution and protein identification from complex proteomic samples. The comparison between the pH gradient SCX-RP 2D HPLC method and traditional salt gradient SCX-RP method was presented. A two-step sample prefractionation method utilizing microwave-assisted tryptic digestion to improve the identification of insoluble proteins was also introduced. This novel 2D HPLC-MS/MS method was applied to the heart proteomic sample of the zebrafish, Danio rerio, to provide comprehensive cardiac proteomic profiling of this important model organism for cardiovascular and environmental toxicology studies.
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