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Updated: Jun 2, 2026

Comprehensive Workflow of Mass Spectrometry-based Shotgun Proteomics of Tissue Samples
Published on: November 13, 2021
Characterizing the proteome of the human dorsolateral prefrontal cortex by shotgun mass spectrometry
Daniel Martins-de-Souza1, Paul C Guest, Hannah Steeb
1Institute of Biotechnology, University of Cambridge, Cambridge, UK. dm533@cam.ac.uk
Abstract:
The studies of neuropsychiatric disorders would be facilitated by enhanced knowledge of the dorsolateral prefrontal cortex (DLPFC) proteome. To construct a data set of human DLPFC proteins, protein extracts were prepared from 12 postmortem brains focussing on the DLPFC region (Brodmann area 9) and analyzed using a combined gel electrophoresis and shotgun mass spectrometry approach, featuring data-independent label-free nanoflow liquid chromatography mass spectrometry (nLC-MS(E)). The detected mass/time features were aligned and annotated using the results from ProteinLynx Global Server. The resulting data set comprised 488 unique and accurately identified proteins, with stringent identification by a minimum of two distinct peptides detected at least in >75% of samples. These proteins were involved predominantly in cytoskeletal architecture, metabolism, transcription/translation, and synaptic function. Combination of this data set with that obtained by our previous characterization of the same brain region results in a total of 755 unique proteins, making this the most comprehensive analysis of this important brain region to date.
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