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J.Health Sci., 49(2), 156-159, 2003
Epitope-Tagged Metallothionein: Its Biochemical and
Immunocytochemical Applications
Masami Ishido* and Junko Sayato-Suzuki
Division of Environmental Health Sciences National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba 305-8506, Japan
We developed epitope-tagged metallothionein IIA cDNA by the addition of an 8-amino acid sequence,
DYKDDDDK, at the N-terminus metallothionein using polymerase chain reaction. The fusion proteins were
expressed by transfection in human embryonic kidney 293 (HEK-293) cells and were specifically recognized by
commercially available antibody, an anti-Flag M2 monoclonal antibody. Furthermore, the fusion protein was
detectable on a Western blot, whose migration was expectedly at 7000-8000 in an SDS-denatured form. Thus,
availability of the epitope-tagged metallothionein will facilitate the investigation of cellular mechanisms of the
protein such as its nuclear localization mechanism.
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