[The expression and antibody preparation of S100A10 protein].
OBJECTIVE
To prepare S100A10 protein and its specific polyclonal antibody.


METHODS
The full-length gene fragment of S100A10 was amplified by PCR, and then cloned into pET28a(+) prokaryotic expression vector. After transformation, the vector was induced to express the recombinant (S100A10)(2) protein by IPTG in E.coli BL21 (DE3). The recombinant (S100A10)(2) was then purified by Ni-NTA resin. (S100A10)(2)-specific polyclonal antibody was prepared using the purified recombinant (S100A10)(2) protein as antigen to inoculate rabbit intradermally. The title and specificity of the polyclonal antibody were determined by ELISA and Western blotting.


RESULTS
The study successfully constructed the prokaryotic recombinant expression vector pET28a(+)-(S100A10)(2), and obtained the purified recombinant (S100A10)(2) protein and polyclonal antibody with high titer and specificity.


CONCLUSIONS
The prokaryotic expression and purification system for S100A10 has been established and polyclonal antibody of (S100A10)(2) been prepared, which provides helpful fools for further researches on S100A10.
Institute of Molecular and Biology, Three Gorges University, Yichang 443002; Department of Obstetrics and Gynecology, People's Hospital of China Three Gorges University, Yichang 443000, China.
Dandan
D
Xin
Institute of Molecular and Biology, Three Gorges University, Yichang 443002, China.
Xiaohua
X
Zou
Department of Obstetrics and Gynecology, People's Hospital of China Three Gorges University, Yichang 443000, China.
Manzhen
M
Zuo
Institute of Molecular and Biology, Three Gorges University, Yichang 443002, China.
Chaoqi
C
Liu
D000818:Animals; D017306:Annexin A2 / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D010957:Plasmids; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis*; D009418:S100 Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D000099248:S100 Calcium Binding Protein A10
D017306:Annexin A2; D000906:Antibodies; D011994:Recombinant Proteins; D009418:S100 Proteins; D000099248:S100 Calcium Binding Protein A10; C472822:ANXA2 protein, human