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Related Experiment Video

Updated: Jul 12, 2026

Single-Cell Proteomics Preparation for Mass Spectrometry Analysis Using Freeze-Heat Lysis and an Isobaric Carrier
06:13

Single-Cell Proteomics Preparation for Mass Spectrometry Analysis Using Freeze-Heat Lysis and an Isobaric Carrier

Published on: December 9, 2022

Improved Surfactant-Assisted One-Pot Sample Preparation for Robust Convenient Single-Cell Proteomics.

Reta Birhanu Kitata1, Zhangyang Xu1, Nadia Bayou2

  • 1Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99354, United States.

Journal of Proteome Research
|July 10, 2026
PubMed
Summary

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A new single-cell proteomics (SCP) method, iSOP-MS, uses a 3 μL volume for robust and convenient analysis. This improved surfactant-assisted one-pot processing allows for cost-effective, quantitative SCP with high protein group detection from single cells.

Area of Science:

  • Proteomics
  • Biochemistry
  • Analytical Chemistry

Background:

  • Current single-cell proteomics (SCP) methods often require specialized devices or extensive measures to prevent evaporation, limiting accessibility and robustness.
  • Sub-microliter to 1-microliter processing volumes are common, posing challenges for routine laboratory use.

Purpose of the Study:

  • To develop a robust, convenient, and cost-effective single-cell proteomics method using a standard 384-well plate format.
  • To improve the accessibility and analytical robustness of SCP technologies.

Main Methods:

  • Developed iSOP (improved surfactant-assisted one-pot processing) using a 3 μL processing volume in a sealed 384-well plate.
  • Optimized enzyme mixture (2 ng each of trypsin and Lys-C) for enhanced sensitivity and robustness.
Keywords:
384-well platecellular heterogeneityiSOP-MSlow μL processing volumesingle-cell proteomics (SCP)

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

Published on: February 7, 2022

Related Experiment Videos

Last Updated: Jul 12, 2026

Single-Cell Proteomics Preparation for Mass Spectrometry Analysis Using Freeze-Heat Lysis and an Isobaric Carrier
06:13

Single-Cell Proteomics Preparation for Mass Spectrometry Analysis Using Freeze-Heat Lysis and an Isobaric Carrier

Published on: December 9, 2022

Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry
11:12

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

Published on: February 7, 2022

  • Utilized a standard liquid chromatography-mass spectrometry (LC-MS) platform for analysis.

    • Main Results:

    • iSOP-MS reliably detected and quantified approximately 1200-1800 protein groups from single HeLa or MCF7 cells.
    • Analysis of neuroblastoma cell lines (BE2-C and SK-N-SH) identified an average of ~1700 and ~2050 protein groups per cell, respectively.
    • Demonstrated precise characterization of cellular heterogeneity using the iSOP-MS method.

    Conclusions:

    • iSOP-MS offers a more robust and convenient alternative for routine, cost-effective, quantitative single-cell proteomics.
    • The method overcomes limitations of existing SCP approaches, enhancing broad accessibility.
    • Enables detailed cellular heterogeneity analysis through high-throughput proteomic profiling.