Optimization of proteomics sample preparation for identification of host and bacterial proteins in mouse feces.

ABSTRACT: Bottom-up proteomics is a powerful method for the functional characterization of mouse gut microbiota. To date, most of the bottom-up proteomics studies of the mouse gut rely on limited amounts of fecal samples. With mass-limited samples, the performance of such analyses is highly dependent on the protein extraction protocols and contaminant removal strategies. Here, protein extraction protocols (using different lysis buffers) and contaminant removal strategies (using different types of filters and beads) were systematically evaluated to maximize quantitative reproducibility and the number of identified proteins. Overall, our results recommend a protein extraction method using a combination of sodium dodecyl sulfate (SDS) and urea in Tris-HCl to yield the greatest number of protein identif

SUBMITTER: Baniasad M 

PROVIDER: S-EPMC9393048 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature