Phos-tag SDS-PAGE database casein ovalbumin

Image

Figs shows the effect of Mn²⁺-Phos-tag on the mobility of phosphorylated proteins (α-casein, β-casein, and ovalbumin) in SDS-PAGE . In the absence of Mn²⁺-Phos-tag (i.e. normal SDSPAGE),
the mobilities of phosphorylated and dephosphorylated proteins are almost the same. In the presence of Mn²⁺-Phos-tag (50, 100, and 150 M), a difference in mobility between the phosphorylated and the corresponding dephosphorylated proteins is observed. The Phos-tag ligand without Mn²⁺ did not show any mobility shifts of phosphorylated proteins (data not shown). The Rf values of all samples become smaller dose-dependently. This is possibly due to electrostatic interaction between cationic Mn²⁺-Phos-tag and an-ionic SDS-bound proteins. Interestingly β-casein (5P) appears as eight bands using 100 M Mn²⁺-Phos-tag, indicating
the existence of at least eight isotypes with a different number of phosphomonoester dianions.

Reference	Kinoshita, E., Kinoshita-Kikuta, E., Takiyama, K., and Koike, T. Phosphate-binding tag, a new tool to visualize phosphorylated proteins. Molecular & Cellular Proteomics, 5, 749-757 (2006) DOI:10.1074/mcp.T500024-MCP200
Composition of gel	Mn²⁺–Phos-tag:　0-100µM Polyacrylamide: 10%(w/v)
Detection	CBB stain

Reference

Kinoshita, E., Kinoshita-Kikuta, E., Takiyama, K., and Koike, T.
Phosphate-binding tag, a new tool to visualize phosphorylated proteins. Molecular & Cellular Proteomics, 5, 749-757 (2006)

DOI:10.1074/mcp.T500024-MCP200

Composition of gel

Mn²⁺–Phos-tag:　0-100µM
Polyacrylamide: 10%(w/v)

Detection

CBB stain

α-casein, β-casein

ovalbumin

Image

Related data