Effect of the metal in the reaction between metallothionein and antimetallothionein antibody

1. An antimetallothionein antibody, raised against Cd-carrying metallothionein, was applied in Western blotting of metallothionein. 2. Treatment of the electroblotted nitrocellulose sheets with metals belonging to the periodic system transition groups Ib and IIb, or with Pb, Ni or Cr, considerably enhanced binding of anti-metallothionein. A similar effect was found when the electroblotted sheets were treated with the strong alkylator N-ethylmaleimide. 3. It seems that the binding of metal to metallothionein modifies the configuration of the antibody binding sites by the formation of metal thiolate complexes. 4. Metal treatment of the nitrocellulose sheets after electroblotting, but before application of the primary antibody, offers a convenient method for use in Western blotting to significantly potentiate the reaction between metallothionein and the antimetallothionein antibody.