HPH fractionation was performed on mitochondrial digested peptides after SEC fractionation using Phenomenex Gemini C18 column on an Agilent 1260 Infinity II UPLC system. A 85 min gradient was applied and 42 fractions were collected and dried under SpeedVac.
Gemini c18
Manufactured by Agilent Technologies
The Gemini C18 is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. It features a spherical silica-based stationary phase with chemically bonded C18 alkyl chains, providing high-resolution separations and excellent peak shapes.
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2 protocols using gemini c18
1
Mitochondrial Peptide Fractionation
2
HPLC Purity Analysis of PAs
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The purity of the PAs after HPLC purification was determined by analytical LC-MS using an Agilent 1200 system equipped with a Phenomenex Gemini C18, 1 × 100 mm column, detector Agilent 6520 Q-TOF LC-MS. Gradient: acetonitrile 5% for 5 min at 50 µL min−1, 5–95% over 25 min at 50 µL min−1 followed by 95% for 5 min at µL min−1. Peaks were detected at 220 nm.
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