Identification of metallothionein isoforms on capillary zone electrophoresis by adding anti-metallothionein antibody.
The aim of this study was to identify metallothionein (MT) isoforms in mouse liver by using capillary zone electrophoresis (CZE). Purified MT-1 and MT-2 isoforms were completely separated by CZE using a polyacrylamide-coated tube at physiologic pH. There were two peaks in the cytosol fraction prepared from zinc-injected mouse liver, in which the migration times corresponded with those of purified MT-1 and MT-2 isoforms. When anti-MT monoclonal antibody was added with the purified MT-1 or MT-2 solution, the peaks decreased. Furthermore, the two peaks in the cytosol prepared from Zn-injected mouse liver decreased in a time-dependent manner from the electropherogram after the addition of the antibody. Therefore, those peaks were identified as MT-1 and MT-2 isoforms, respectively. In conclusion, the addition of anti-MT monoclonal antibody to the cytosol fraction of tissues is an effective method for identification of MT isoforms after separation using CZE.
Kubo
K
K
Faculty of Pharmaceutical Sciences, Kinki University, Higashi-Osaka, Osaka, Japan.
Sakita
Y
Y
Okazaki
Y
Y
Otaki
N
N
Kimura
M
M
Minami
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D003600:Cytosol / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D008668:Metallothionein / Q000032:analysis* / Q000378:metabolism; D051379:Mice; D015032:Zinc / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008668:Metallothionein; D015032:Zinc