StageTip-based HAMMOC, an efficient and inexpensive phosphopeptide enrichment method for plant shotgun phosphoproteomics

Methods Mol Biol. 2014;1072:595-607. doi: 10.1007/978-1-62703-631-3_40.

Abstract

Phosphopeptide enrichment is the most critical step for successful LC-MS/MS-based shotgun phosphoproteomics. Recent technological improvements have made selective phosphopeptide enrichment from non-fractionated whole cell lysate digests with a single-step procedure possible. Here, a handy protocol is described for phosphopeptide enrichment from plant materials using hydroxy acid-modified metal oxide chromatography (HAMMMOC) with a stop-and-go-extraction tip (StageTip).

MeSH terms

  • Chromatography / instrumentation*
  • Chromatography / methods*
  • Phosphopeptides / isolation & purification*
  • Phosphoproteins / isolation & purification*
  • Plant Proteins / isolation & purification*
  • Proteomics / instrumentation*
  • Proteomics / methods*

Substances

  • Phosphopeptides
  • Phosphoproteins
  • Plant Proteins