Proteomics of mouse liver microsomes: Performance of different protein separation workflows for LC‐MS/MS

Abstract

The mouse liver microsome proteome was investigated using ion trap MS combined with three separation workflows including SDS-PAGE followed by reverse-phase LC of in-gel protein digestions (519 proteins identified); 2-D LC of protein digestion (1410 proteins); whole protein separation on mRP heat-stable column followed by 2-D LC of protein digestions from each fraction (3-D LC; 3703 proteins). The higher number of proteins identified in the workflow corresponded to the lesser percentage of run-to-run reproducibility. Gel-based method yielded a number of predicted membrane proteins similar to LC-based workflows.