Epitope-Tagged Metallothionein: Its Biochemical and Immunocytochemical Applications

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 recogn...

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Bibliographic Details
Published inJournal of Health Science Vol. 49; no. 2; pp. 156 - 159
Main Authors Ishido, Masami, Sayato-Suzuki, Junko
Format Journal Article
LanguageEnglish
Published The Pharmaceutical Society of Japan 2003
Pharmaceutical Society of Japan
Subjects
cellular localization
epitope-tag
metallothionein
Western botting
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Summary: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.
ISSN:1344-9702
1347-5207
DOI:10.1248/jhs.49.156