Section 27
Chapter 26,029

Two-dimensional phosphate-affinity gel electrophoresis for the analysis of phosphoprotein isotypes

Kinoshita, E.; Kinoshita-Kikuta, E.; Matsubara, M.; Aoki, Y.; Ohie, S.; Mouri, Y.; Koike, T.

Electrophoresis 30(3): 550-559


ISSN/ISBN: 0173-0835
PMID: 19156764
DOI: 10.1002/elps.200800386
Accession: 026028107

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Herein, we describe three kinds of 2-DE using phosphate-affinity PAGE for the analysis of phosphoprotein isotypes. The first dimension is a urea-PAGE, IEF/NEPHGE, or SDS-PAGE, which are widely used. The second dimension is a phosphate-affinity SDS-PAGE using a phosphate-binding tag molecule, Phos-tag (Mn(2+)-Phos-tag SDS-PAGE). The first 2-D procedure coupling urea-PAGE and Mn(2+)-Phos-tag SDS-PAGE was applied to the separation of beta-casein phosphoisotypes.

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