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...

Full description

Saved in:
Bibliographic Details
Published in:Journal of Health Science 2003, Vol.49(2), pp.156-159
Main Authors: Ishido, Masami, Sayato-Suzuki, Junko
Format: Article
Language:English
Subjects:
cellular localization
epitope-tag
metallothionein
Western botting
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
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