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 zebra fish, Danio rerio , to provide comprehensive cardiac proteomic profiling of this important model organism for cardiovascular and environmental toxicology studies. © Springer Science+Business Media New York 2013.
CITATION STYLE
Zhang, J., Lanham, K. A., Heideman, W., Peterson, R. E., & Li, L. (2013). Characterization of zebra fish cardiac proteome using online ph gradient SCX-RP HPLC-MS/MS platform. Methods in Molecular Biology, 1005, 119–127. https://doi.org/10.1007/978-1-62703-386-2_10
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