Figure 3.

Optimized workflow for proteomic analysis of pancreatic fluid. Pancreatic fluid is collected via ePFT. After particulates are removed by centrifugation, proteins are extracted from the remaining supernatant with TCA. The protein pellet is reconstituted in Laemmli buffer, alkylated, and reduced prior to analysis by SDS-PAGE. GeLC-MS/MS analysis is performed, in which gel lanes are divided into smaller segments, which are individually in-gel tryptically digested. Digested peptides are eluted from a nanoflow C18 reversed-phased column into a mass spectrometer for accurate mass analysis. The resulting mass spectra are processed to determine the peptides and eventually the proteins from which these peptides originate.