title: Improved Proteome and Phosphoproteome Analysis on a Cation Exchanger by a Combined Acid and Salt Gradient.
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.
authors.affiliation: Laboratory of Proteome Research, National Institute of Biomedical Innovation, Health and Nutrition , Ibaraki, Osaka 567-0085, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Adachi
authors.affiliation: Laboratory of Proteome Research, National Institute of Biomedical Innovation, Health and Nutrition , Ibaraki, Osaka 567-0085, Japan.
authors.forename: Kazunari
authors.initials: K
authors.lastname: Hashiguchi
authors.affiliation: Laboratory of Proteome Research, National Institute of Biomedical Innovation, Health and Nutrition , Ibaraki, Osaka 567-0085, Japan.
authors.forename: Maiko
authors.initials: M
authors.lastname: Nagano
authors.affiliation: Laboratory of Proteome Research, National Institute of Biomedical Innovation, Health and Nutrition , Ibaraki, Osaka 567-0085, Japan.
authors.forename: Misako
authors.initials: M
authors.lastname: Sato
authors.affiliation: Laboratory of Proteome Research, National Institute of Biomedical Innovation, Health and Nutrition , Ibaraki, Osaka 567-0085, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Sato
authors.affiliation: Laboratory of Proteome Research, National Institute of Biomedical Innovation, Health and Nutrition , Ibaraki, Osaka 567-0085, Japan.
authors.forename: Kazuna
authors.initials: K
authors.lastname: Fukamizu
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University , Kyoto 606-8501, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Ishihama
authors.affiliation: Laboratory of Proteome Research, National Institute of Biomedical Innovation, Health and Nutrition , Ibaraki, Osaka 567-0085, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tomonaga
mesh_terms: D002412:Cations / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D020543:Proteome; D012492:Salts / Q000737:chemistry
chemical_list: D002412:Cations; D010455:Peptides; D020543:Proteome; D012492:Salts