DIGE-Based Phosphoproteomic Analysis

Taras Stasyk; Lukas Alfons Huber

doi:10.1007/978-1-4939-7268-5_7

DIGE-Based Phosphoproteomic Analysis

Methods Mol Biol. 2018:1664:79-86. doi: 10.1007/978-1-4939-7268-5_7.

Authors

Taras Stasyk¹, Lukas Alfons Huber²

Affiliations

¹ Division of Cell Biology, Biocenter, Innsbruck Medical University, Innrain 80-82, A-6020, Innsbruck, Austria. taras.stasyk@i-med.ac.at.
² Division of Cell Biology, Biocenter, Innsbruck Medical University, Innrain 80-82, A-6020, Innsbruck, Austria.

PMID: 29019126
DOI: 10.1007/978-1-4939-7268-5_7

Abstract

Here, we describe the detailed step-by-step protocol for detection of phosphoproteins in two-dimensional difference gel electrophoresis (DIGE) gels. A standard DIGE protocol is combined with subsequent post-staining with phosphospecific fluorescent dye. The combination of these two methods complements 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

Image Processing, Computer-Assisted
Phosphoproteins*
Proteome
Proteomics* / methods
Two-Dimensional Difference Gel Electrophoresis* / methods

Substances

Phosphoproteins
Proteome