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Related Experiment Video

Updated: May 12, 2026

A Protein Suspension-Trapping Sample Preparation for Tear Proteomics by Liquid Chromatography-Tandem Mass Spectrometry
08:20

A Protein Suspension-Trapping Sample Preparation for Tear Proteomics by Liquid Chromatography-Tandem Mass Spectrometry

Published on: December 1, 2023

A Suspension-Trapping Protocol for Bottom-Up Proteomics Sample Preparation.

Joseph Schrader1, Dennis Province2, Nicholas A DaSilva3

  • 1Department of Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, RI, USA.

Bio-Protocol
|May 11, 2026
PubMed
Summary

This study presents an optimized Suspension Trapping (S-Trap) workflow for bottom-up proteomics sample preparation. The method efficiently processes challenging biofluids like plasma, serum, and cerebrospinal fluid (CSF), improving reproducibility and accessibility.

Keywords:
LC-MSMass spectrometryProteomicsS-trapSample preparation

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Last Updated: May 12, 2026

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Published on: December 1, 2023

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Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry

Published on: February 7, 2022

Area of Science:

  • Proteomics
  • Biochemistry
  • Analytical Chemistry

Background:

  • Sample preparation is critical for reproducible bottom-up proteomics.
  • Existing methods like in-gel, in-solution, and filter-aided digestion have limitations.
  • Sodium dodecyl sulfate (SDS) can interfere with mass spectrometry analysis.

Purpose of the Study:

  • To detail an optimized Suspension Trapping (S-Trap) workflow for biofluid proteomics.
  • To provide protocols for both manual and high-throughput sample processing.
  • To improve consistency and accessibility of S-Trap for challenging samples.

Main Methods:

  • Developed an S-Trap protocol for plasma, serum, and cerebrospinal fluid (CSF).
  • Included optional high-abundance protein depletion.
  • Offered manual tube-based and 96-well plate formats.
  • Integrated reduction, alkylation, digestion, and peptide elution steps.

Main Results:

  • The S-Trap method efficiently captures and digests proteins.
  • It effectively removes interferents like SDS.
  • The workflow is optimized for low protein content and high lipid samples.
  • Achieved high peptide recovery and improved reproducibility.

Conclusions:

  • The S-Trap workflow enhances biofluid proteomics sample preparation.
  • It offers flexibility with manual and high-throughput options.
  • This method facilitates robust and reproducible proteomic discoveries from challenging samples.