Figure 1.
Workflow of standard-scale and microgram-scale of TMT-LC/LC-MS/MS. (A) Standard-scale pipeline. Digested proteins (peptides) are desalted and labeled by TMT (TMT/peptide ratio of 1.5:1). The pooled samples are then desalted and analyzed by LC/LC-MS/MS. (B) Microgram-scale pipeline. Proteins in the tissue lysate are digested into peptides with excess trypsin to shorten digestion time. Without desalting, the digested tissue lysate is directly labeled by TMT (TMT/tissue ratio of 3:1). The pooled sample is analyzed by LC/LC-MS/MS with highly sensitive LC columns.