Analysis of Protein Phosphorylation Using Phos‐Tag Gels. (25th July 2018)

• Record Type: Journal Article
• Authors: Nagy, Zoltan; Comer, Shane; Smolenski, Albert
• Editors: Coligan, John E.; Dunn, Ben M.; Speicher, David W.; Wingfield, Paul T.
• Is Part Of: Current protocols in protein science. Volume 93(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-07-25
• Subjects: Phos‐tag -- phosphorylation -- phosphate‐affinity electrophoresis -- protein kinase; Proteins -- Laboratory manuals; Proteins; Clinical Laboratory Techniques; Genetic Techniques; Immunologic Techniques; Proteins; Laboratory manuals; 572.6028
• DOI: 10.1002/cpps.64
• Languages: English
• ISSNs: 1934-3655

Abstract

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 Zn 2+ ‐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.