PTM-Works » Quantitative Histone Profiling

Quantitative Histone Profiling

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Overview

This service enables a quantitative comparison of two samples for global histone marks. Histones are isolated from cells or tissue followed by quantitative mass spectrometry analysis in a single pairwise experiment using the “One-Pot” approach* with light and heavy propionylation.

* One-Pot Quantitative Mass Spectrometry Characterization of Histones”, Garcia et al., J. Proteome Research, 8, 5367-5374(2009).

Steps Involved

  • Isolation of histones from cells or tissue using the Active Motif Histone Purification Kit. Submission of 10million cells or 25mg of tissue (or greater) is required.
  • Block lysine side chains using d0 propionic anhydride.
  • Solution digestion with trypsin.
  • Label peptide N-terminii with either d0 (sample 1) or d10 (sample 2) propionic anhydride.
  • Mix samples 1:1.
  • 90min LC/MS/MS in analytical duplicate on Orbitrap mass spectrometer.
  • Peak integration for over 70 peptide/PTM combinations across H2A, H2B, H3 and H4.
  • Differential analysis.
  • Report in PDF and Excel.

Deliverables

  • Written report containing details of your samples, our methods and a summary of the data.
  • An Excel workbook containing the detailed quantitative data.