[Expression and purification of human MT-2a fusion protein in prokaryotic cells and preparation of its antiserum].
OBJECTIVE
To express and purify human MT-2a in prokaryotic cells and to prepare the MT-2a-specific rabbit antiserum.


METHODS
GST-MT-2a fusion protein was expressed after IPTG induction and further purified with Glutathione Sepharose 4B. Then the purified GST-MT-2a fusion protein was used to immunize New Zealand rabbits. The titer and specificity of rabbit antiserum were evaluated by double immunodiffusion, ELISA and Western blot.


RESULTS
GST-MT-2a fusion protein was highly expressed. The final yield of the pure GST-MT-2a was about 38 mg per liter of bacterial culture. Its antiserum with high specificity and potency was also obtained.


CONCLUSIONS
The successful expression of GST-MT-2a fusion protein in E. coli and the preparation of MT-2a specific rabbit antiserum will be valuable for the study on the function of human MT-2a.
Zhou
Min
M
Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha 410078, China. hezhimin2005@yahoo.com
Yang
Fang
F
He
Zhi-min
ZM
Liu
Xiao-rong
XR
Chen
Zhu-chu
ZC
Sun
Lin
L
Sun
Yu
Y
D000918:Antibody Specificity; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D008668:Metallothionein / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D011387:Prokaryotic Cells / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D012995:Solubility
D007106:Immune Sera; C491053:MT2A protein, human; D011993:Recombinant Fusion Proteins; D008668:Metallothionein