Open AccessProduction of the Polyclonal Anti‐human Metallothionein 2A Antibody with Recombinant Protein Technology
Author(s) -
MARIKAR Faiz M. M. T.,
SUN QiMing,
HUA ZiChun
Publication year - 2006
Publication title -
acta biochimica et biophysica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.771
H-Index - 57
eISSN - 1745-7270
pISSN - 1672-9145
DOI - 10.1111/j.1745-7270.2006.00167.x
Subject(s) - polyclonal antibodies , recombinant dna , fusion protein , antiserum , microbiology and biotechnology , western blot , gel electrophoresis , biology , polyacrylamide gel electrophoresis , protein a/g , antibody , protein g , chemistry , biochemistry , enzyme , gene , immunology
Metallothionein 2A (MT2A) is a small stress response protein that can be induced by exposure to toxic metals. It is highly expressed in breast cancer cells. In this study, the cDNA encoding the human MT2A protein was expressed as glutathione S‐transferase (GST) fusion protein in Escherichia coli. Recombinant MT2A proteins were loaded onto 12% sodium dodecylsulfate‐polyacrylamide gel and separated by electrophoresis, the recombinant protein was visualized by Coomassie blue staining and the 33 kDa recombinant GST‐MT2A fusion protein band was cut out from the gel. The gel slice was minced and used to generate polyclonal antisera. Immunization of rabbit against MT2A protein allowed the production of high titer polyclonal antiserum. This new polyclonal antibody recognized recombinant MT2A protein in Western blot analysis. This low‐cost antibody will be useful for detection in various immuno‐assays. Edited by
Ming‐Hua XU