DIGE-Based Phosphoproteomic Analysis

Taras Stasyk; Lukas Alfons Huber

doi:10.1007/978-1-0716-2831-7_7

DIGE-Based Phosphoproteomic Analysis

Methods Mol Biol. 2023:2596:97-104. doi: 10.1007/978-1-0716-2831-7_7.

Authors

Taras Stasyk¹, Lukas Alfons Huber²

Affiliations

¹ Institute of Cell Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria. taras.stasyk@i-med.ac.at.
² Institute of Cell Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria.

PMID: 36378433
DOI: 10.1007/978-1-0716-2831-7_7

Abstract

Here, we describe a detailed step-by-step protocol for the detection of phosphoproteins in two-dimensional difference gel electrophoresis (2D-DIGE) gels. A standard 2D-DIGE protocol is combined with subsequent post-staining with phosphospecific fluorescent dye. The combination of these two methods complements 2D-DIGE-based proteome profiling by fluorescence detection of phosphoproteins in the same gel providing additional possibility for sensitive and accurate quantification of the differentially regulated phosphoproteins in biological samples.

Keywords: 2D-DIGE; Phosphoprotein; Phosphoproteomics.

MeSH terms

Electrophoresis, Gel, Two-Dimensional / methods
Fluorescent Dyes
Phosphoproteins*
Proteome
Proteomics* / methods
Staining and Labeling
Two-Dimensional Difference Gel Electrophoresis / methods

Substances

Phosphoproteins
Fluorescent Dyes
Proteome