Detection of Multisite Phosphorylation of Intrinsically Disordered Proteins Using Phos-tag SDS-PAGE

• DOI: 10.1007/978-1-0716-0524-0_40
• Publication Date: 2020-07-21T17:04:25Z
• Authors: Mihkel Örd; Mart Loog
• Keywords: Phos

Abstract Phos-tag TM SDS-PAGE is a method that enables electrophoretic separation of proteins based on their phosphorylation status. With SDS-PAGE, it possible to discriminate between different phosphoforms level and the number phosphorylated sites, determine stoichiometry products. useful for analyzing disordered with multiple sites can be used any downstream applications in combination conventional example, Western blotting mass-spectrometry. To obtain best results however, often necessary optimize gel composition. Depending molecular weight phosphoryl groups added protein, composition or running conditions should used. Here, we provide protocols Mn 2+ - Zn -Phos-tag give examples how characteristics behave gels various concentrations.