Elegant English Translation for Peptide Labeling and Fractionation Protocol

The Thermo Scientific TMT Reagent 6-plex Kit facilitates the labeling of peptides. Manufacturer guidelines advocate the use of the TMT reagent to label 100 μg peptide mixtures per sample, followed by the fractionation of the labeled peptides using the Thermo Scientific High pH Reversed-Phase Peptide Fractionation Kit. In essence, the dried peptide mixture undergoes reconstitution and acidification with a 0.1% TFA solution, prior to being loaded onto the equilibrated, high-pH, reversed-phase fractionation spin column. Subsequent to desalting through low-speed centrifugation, the acetonitrile concentrations within the volatile high-pH elution solution are incrementally augmented. The bound peptides are then applied to the columns, resulting in the collection of ten distinct fractions via centrifugation. The collected fractions undergo desalting using C18 Cartridges and are concentrated using vacuum centrifugation.