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  • Title: Detection of metallothionein on nitrocellulose membrane using Western blotting technique and its application to identification of cadmium-binding proteins.
    Author: Aoki Y, Kunimoto M, Shibata Y, Suzuki KT.
    Journal: Anal Biochem; 1986 Aug 15; 157(1):117-22. PubMed ID: 3766954.
    Abstract:
    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.
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