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Related Experiment Video

Updated: Jan 13, 2026

Automated Sample Preparation for the Multiplexed Analysis of Single-Cell Histone Post-Translational Modifications (sc-hPTM2)

Automated Sample Preparation for the Multiplexed Analysis of Single-Cell Histone Post-Translational Modifications (sc-hPTM2)

Published on: December 19, 2025

Automated Sample Preparation for the Multiplexed Analysis of Single-Cell Histone Post-Translational Modifications

Giulia Barotti¹, Ronald Cutler¹, Joshua Cantlon²

¹Albert Einstein College of Medicine.

Journal of Visualized Experiments : Jove

|January 6, 2026

Summary

This summary is machine-generated.

This study introduces an automated single-cell proteomics workflow for analyzing histone post-translational modifications (hPTMs). The method enables sensitive, reproducible investigation of chromatin heterogeneity and epigenetic changes at the single-cell level.

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Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis

Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis

Published on: May 17, 2016

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Last Updated: Jan 13, 2026

Automated Sample Preparation for the Multiplexed Analysis of Single-Cell Histone Post-Translational Modifications (sc-hPTM2)

Automated Sample Preparation for the Multiplexed Analysis of Single-Cell Histone Post-Translational Modifications (sc-hPTM2)

Published on: December 19, 2025

Author Spotlight: Enhanced Histone PTM Isomer Identification Through LC-TIMS-ToF MS/MS and PASEF

Author Spotlight: Enhanced Histone PTM Isomer Identification Through LC-TIMS-ToF MS/MS and PASEF

Published on: January 12, 2024

Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis

Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis

Published on: May 17, 2016

Area of Science:

Epigenetics and Molecular Biology
Proteomics
Cellular Biology

Background:

Histone post-translational modifications (hPTMs) regulate chromatin organization and gene expression.
Dysregulated hPTMs are linked to cancer, aging, and developmental disorders.
Current proteomics methods for hPTMs are typically bulk analyses, lacking single-cell resolution.

Purpose of the Study:

To develop an automated, sensitive workflow for single-cell histone PTM analysis.
To overcome technical challenges of low histone abundance and high modification levels in single cells.
To enable investigation of chromatin heterogeneity and epigenetic perturbations at single-cell resolution.

Main Methods:

An automated nano liquid handling system for nanoliter-scale processing of individual cells.
Digestion using ArgC Ultra protease, avoiding lysine-blocking derivatization.
A two-plex labeling strategy using propionic anhydride and its deuterated analogue for quantitative comparison.

Main Results:

A streamlined and sensitive protocol for single-cell histone PTM analysis was established.
The workflow demonstrated high reproducibility and minimal sample handling.
The method was successfully applied to analyze histone acetylation changes in response to sodium butyrate treatment in hepatocellular carcinoma spheroids.

Conclusions:

The developed workflow provides a powerful tool for high-resolution analysis of histone PTMs.
This advancement facilitates the study of epigenetic heterogeneity within cell populations.
The protocol opens new avenues for understanding the role of hPTMs in various biological processes and diseases.