Resource Summary Report  New Search Previous Search Results

URL: https://dx.doi.org/DOI:10.17504/protocols.io.bfx6jpre

Authors: Jennifer Gin, Yan Chen, Christopher Petzold

Group: LBNL-omics

Summary: Recent improvements in the speed and sensitivity of liquid chromatography-mass spectrometry systems have driven progress toward system-wide characterization of the proteome of many species. These efforts create large proteomic datasets that provide insight into biological processes and identify diagnostic proteins whose abundance changes significantly under different experimental conditions. Consequently, it is important to have reproducible sample preparation methods that consist of mixing, various centrifugation and incubation steps, and an extended tryptic digestion step. We developed a high-throughput sample preparation workflow that consists of cell lysis, protein precipitation, protein resuspension, protein quantification, and normalization of protein concentration followed by standard bottom-up proteomic procedures of reducing and blocking cysteine residues and tryptic digestion.This protocol was adapted from the manual sample preparation method found in Chen, Y., et al. "Automated “Cells-To-Peptides” Sample Preparation Workflow for High-Throughput, Quantitative Proteomic Assays of Microbes."Journal of proteome research 18.10 (2019): 3752-3761.

Affiliations: Lawrence Berkeley National Laboratory, Lawrence Berkeley National Laboratory, Lawrence Berkeley National Laboratory

External URL: https://pubs.acs.org/doi/abs/10.1021/acs.jproteome.9b00455

Version: 1

Publication Date: 2020

Expand All Collapse