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Published on: March 7, 2014
Improved Proteome and Phosphoproteome Analysis on a Cation Exchanger by a Combined Acid and Salt Gradient
Jun Adachi1, Kazunari Hashiguchi1, Maiko Nagano1
1Laboratory of Proteome Research, National Institute of Biomedical Innovation, Health and Nutrition , Ibaraki, Osaka 567-0085, Japan.
Abstract:
Currently used elution methods for strong cation exchange (SCX) chromatography are based on two principles: salt and pH gradient. In this paper, we report the first observation of peptide elution by acid gradient. The degree of peptide separation using C18-SCX StageTip was greatly improved by our acid and salt-based elution method compared with a salt-based elution method. This development enabled us to identify over 22 000 phosphopeptides from 2 mg of protein without labor-intensive sample preparation. Our method is simple, robust, scalable, and low-cost and can be easily implemented without any special equipment or techniques.
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