Work

Analysis of Protein Phosphorylation Using Phos‐Tag Gels

Year: 2018

Type: article

Abstract: Phos‐tag gels are recent tools to dissect protein phosphorylation that operate by inducing a shift in the electrophoretic mobility of phosphorylated proteins compared to their nonphosphorylated counterparts. This article describes the preparation and electrophoresis of Zn2+‐Phos‐tag gels along with electrotransfer of the separated phospho‐ and nonphosphoproteins onto a PVDF membrane using either wet‐tank or semidry transfer. We also discuss the theory behind the technology with critical parameters to keep in mind for its successful application. © 2018 by John Wiley & Sons, Inc.

Source: Current Protocols in Protein Science

DOI: 10.1002/cpps.64

Authors: Zoltan Nagy, Shane Comer, Albert Smolenski

Institutions: University of Birmingham, University College Dublin, Royal College of Surgeons in Ireland

Language: English

Topic: Advanced Proteomics Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: green

Funders: British Heart Foundation, Science Foundation Ireland

Awards: Generation and characterization of a phosphorylation site specific antibody against serine 7 of Rap1GAP2 for application in the diagnosis of platelet reactivity, Regulation of platelet homeostasis and thrombosis by the kinase-phosphatase pair Csk-CD148