Comparison of bottom-up proteomic approaches for LC-MS analysis of complex proteomes.

ABSTRACT: Discovery-based proteomic studies aim to answer important biological questions by identifying as many proteins as possible. In order to accomplish this lofty goal, an effort must be placed on determining an optimal workflow that maximizes protein identifications. In this study, we compare protein extraction, digestion and fractionation methods for bottom-up proteomics using a human colon cancer cell line as our model system. Four different buffers for protein extraction, two digestion approaches, as well as three sample fractionation methods were evaluated in order to determine an accessible workflow that gives maximal protein identifications. Samples comparing these workflows were analyzed via UPLC paired with tandem MS on a Q-Exactive mass spectrometer. Our goal is to determine an optima

SUBMITTER: Weston LA 

PROVIDER: S-EPMC3839868 | biostudies-literature | 2013 Sep

REPOSITORIES: biostudies-literature