Identification of metallothionein in cultured cells by immunoblotting and immunofluorescence using a new monoclonal antibody.
A hybridoma clone (MT45-5-3) producing an IgG-class monoclonal antibody specific for metallothioneins (MTs) was established. The monoclonal antibody (MT45) cross-reacted with mouse, rat and rabbit Cd(2+)-induced MTs 1 and 2 and Zn(2+)-induced MT 2 as assessed by enzyme-linked immunosorbent assay. When the antibody was used to detect the MTs transferred to nylon membranes after SDS-polyacrylamide gel electrophoresis, the antibody reacted with cultured human cell MTs as well as rat, mouse and rabbit MTs 1 and 2 even after carboxymethylation. The antibody could be used for the indirect immunofluorescence test for Cd(2+)-induced MTs in cultured human and mouse cells.
Kikuchi
Y
Y
Department of Microbiology, Hoshi College of Pharmacy, Tokyo, Japan.
Irie
M
M
Ikebuchi
H
H
Sawada
J
J
Terao
T
T
D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D003429:Cross Reactions; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D015151:Immunoblotting / Q000379:methods; D007074:Immunoglobulin G; D008668:Metallothionein / Q000032:analysis* / Q000276:immunology*; D051379:Mice; D011817:Rabbits; D051381:Rats; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008668:Metallothionein