Detection of carboxymethylmetallothionein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
A sensitive method for detecting metallothioneins (MTs) by using silver staining and autoradiography after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of carboxymethylated MTs is described. Carboxymethylation of MTs is indispensable because it prevents their aggregation, thereby allowing each of them to appear as a single band using SDS-PAGE. Metallothioneins can be detected with a limit of nanogram levels per lane. This method can be applied to MTs induced in in vitro cultured cells and in in vivo tissues.
Kimura
M
M
Koizumi
S
S
Otsuka
F
F
D000818:Animals; D001345:Autoradiography / Q000379:methods; D002478:Cells, Cultured; D003545:Cysteine; D003600:Cytosol / Q000737:chemistry; D004229:Dithiothreitol; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007202:Indicators and Reagents; D007461:Iodoacetates; D019807:Iodoacetic Acid; D008099:Liver / Q000737:chemistry; D008668:Metallothionein / Q000031:analogs & derivatives* / Q000032:analysis; D012967:Sodium Dodecyl Sulfate; D013462:Sulfur Radioisotopes
D007202:Indicators and Reagents; D007461:Iodoacetates; D013462:Sulfur Radioisotopes; D012967:Sodium Dodecyl Sulfate; D008668:Metallothionein; D003545:Cysteine; D004229:Dithiothreitol; D019807:Iodoacetic Acid