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This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: M700278-MCP200v1 6/11/2032    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Glossary Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zhang, X. Articles by Roepstorff, P. Search for Related Content PubMed PubMed Citation Articles by Zhang, X. Articles by Roepstorff, P. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 6:2032-2042, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Highly Efficient Phosphopeptide Enrichment by Calcium Phosphate Precipitation Combined with Subsequent IMAC Enrichment^*,S

Xumin Zhang, Juanying Ye, Ole N. Jensen and Peter Roepstorff^¶

From the Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense, Denmark

A new method for enrichment of phosphopeptides in complex mixtures derived by proteolytic digestion of biological samples has been developed. The method is based on calcium phosphate precipitation of the phosphopeptides prior to further enrichment with established affinity enrichment methods. Calcium phosphate precipitation combined with phosphopeptide enrichment using Fe(III) IMAC provided highly selective enrichment of phosphopeptides. Application of the method to a complex peptide sample derived from rice embryo resulted in more than 90% phosphopeptides in the enriched sample as determined by mass spectrometry. Introduction of a two-step IMAC enrichment procedure after calcium phosphate precipitation resulted in observation of an increased number of phosphopeptides.

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				K. Blackburn and M. B. Goshe Challenges and strategies for targeted phosphorylation site identification and quantification using mass spectrometry analysis Brief Funct Genomic Proteomic, December 24, 2008; (2008) eln051v1. [Abstract] [Full Text] [PDF]