Proteomics

Dataset Information

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Highly sensitive phosphoproteomics by tailoring solid-phase extraction to electrostatic repulsion-hydrophilic interaction chromatography

ABSTRACT: The dataset contains the data of key experiments for developing a novel and highly sensitive bottom-up phosphoproteomics strategy termed ERLIC-SCX/RP-LC-MS. We compared two SPE types (RP, SCX) for an ERLIC run with 7 fractions. The results were used to design a workflow for highly sensitive bottom-up phosphoproteomics (third experiment) combing ERLIC in 21 fractions with the fractions 1-9 cleaned by SCX-SPE and the fractions 10-21 by RP-SPE. Up to 50% of the fractions were subjected to LC-MS/MS analysis with 45 h of total MS time per replicate.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Stefan Loroch  

LAB HEAD: Albert Sickmann

PROVIDER: PXD001381 | Pride | 2014-12-02

REPOSITORIES: Pride

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Publications

Highly sensitive phosphoproteomics by tailoring solid-phase extraction to electrostatic repulsion-hydrophilic interaction chromatography.

Loroch Stefan S   Zahedi René Peiman RP   Sickmann Albert A  

Analytical chemistry 20150113 3

In the past decade, several strategies for comprehensive phosphoproteome analysis have been introduced. Most of them combine different phosphopeptide enrichment techniques and require starting material in the milligram range, as a consequence of their insufficient sensitivity. This limitation impairs the applicability of phosphoproteomics to a wide variety of clinical research, where sample material is highly limited. Here we introduce a highly sensitive and easy-to-establish 2D bottom-up strate  ...[more]