Detection of metallothionein on nitrocellulose membrane using Western blotting technique and its application to identification of cadmium-binding proteins.
A convenient method for detecting metallothionein (MT) and other cadmium-binding proteins (Cd-BPs) in tissue cytosol was established. A sample that contains MT was separated on a sodium dodecyl sulfate-polyacrylamide gel and then electrophoretically transferred to a nitrocellulose membrane after reduction with 2-mercaptoethanol (2-ME) on the gel. The membrane was incubated in a buffer containing 109Cd and then subjected to autoradiography. MT-I and -II in rat liver cytosol were detected as radioactive bands together with a Cd-BP of 60,000 Da. On the other hand, three Cd-BPs of 40,000, 29,000, and 24,000 Da were detected in the cytosol when the reduction with 2-ME was omitted.
D000818:Animals; D001345:Autoradiography; D003101:Collodion; D003600:Cytosol / Q000032:analysis; D004586:Electrophoresis; D008099:Liver / Q000032:analysis; D008623:Mercaptoethanol; D008668:Metallothionein / Q000032:analysis*; D008970:Molecular Weight; D051381:Rats
C004203:cadmium-binding protein; D008623:Mercaptoethanol; D003101:Collodion; D008668:Metallothionein