Meat Proteome Profiling Using TMT

The meat samples were prepared following our previous published protocols [5 (link)]. In brief, 1.5 mL of protein lysis buffer (8 M urea, 1% SDS) was used to suspend the powder of meat samples (100 mg). Then, after being incubated on ice for 30 min and sonicated for 2 min, the supernatant was collected and quantified through a BCA Protein Assay Kit (Thermo Scientific). Lastly, 9 pools (5 samples per pool and 3 pools per age stage) with the same concentration were obtained according to protein content. After being digested at 37 °C overnight with trypsin (the mass ratio of trypsin-to-protein at 1:50), the peptides were labeled with 10-plex TMT reagents (Thermo Fisher Scientific) and multiplex labeled samples were obtained.