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Strong cation exchange chromatography (SCX) and LC-ESI-MS/MS analysis

HW Huang-Joe Wang
SC Sung-Fang Chen
WL Wan-Yu Lo
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Peptide separation was performed using a Polysulfoethyl A Column (200 mm L × 2.1 mm i.d., 5 μm, 300 Å, PolyLC, Columbia, MD) with an Agilent 1100 binary HPLC (Agilent Technologies, Wilmington, DE) over a 90 min gradient. Eighty fractions were collected after SCX fractionation and then pooled into 24 fractions. The fractions were purified using C-18 Spin columns (Thermo Scientific, San Jose, CA) for further nano-LC-ESI-MS/MS analysis14.

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