Reprosil pur 120 c18 aq material

The desalted peptides were dissolved in 200 mM HEPES (pH 8.5). Then, the resuspended digested peptides were mixed with TMT reagent dissolved in 100% ACN for 1 h. The labeling reaction was stopped by adding 5% hydroxylamine for 15 min, followed by acidification with TFA. All the peptides labeled with each multiplexed TMT reagent were mixed into the same tube and the mixture was diluted to decrease the concentration of ACN to less than 5%. The TMT-labeled peptides were desalted on SDB-XC StageTips. The information on the peptide amount in each TMT channel for all experiments is shown in Table S3. Note that the peptide amount for each TMT channel was quantified by means of nanoLC-UV at 210 nm using a Thermo Ultimate 3000 RSLCnano system (Germering), an MU701 UV detector (GL Sciences), and a C18 analytical column (150 mm length × 100 μm ID) packed with Reprosil-Pur 120 C18-AQ material (3 μm, Dr. Maisch).