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