Proteomics

Dataset Information

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Defeating major contaminants in Fe3+-IMAC phosphopeptide enrichment

ABSTRACT: Optimized sample preparation workflow for Fe-IMAC phosphopeptide enrichment.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human) Escherichia Coli

SUBMITTER: Simone Lemeer  

LAB HEAD: Simone Lemeer

PROVIDER: PXD008289 | Pride | 2018-02-19

REPOSITORIES: Pride

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Publications

Defeating Major Contaminants in Fe<sup>3+</sup>- Immobilized Metal Ion Affinity Chromatography (IMAC) Phosphopeptide Enrichment.

Potel Clement M CM   Lin Miao-Hsia MH   Heck Albert J R AJR   Lemeer Simone S  

Molecular & cellular proteomics : MCP 20180215 5

Here we demonstrate that biomolecular contaminants, such as nucleic acid molecules, can seriously interfere with immobilized metal ion affinity chromatography (IMAC)-based phosphopeptide enrichments. We address and largely solve this issue, developing a robust protocol implementing methanol/chloroform protein precipitation and enzymatic digestion using benzonase, which degrades all forms of DNA and RNA, before IMAC-column loading. This simple procedure resulted in a drastic increase of enrichmen  ...[more]