[Recombinant expression of human fibrogenic factors and preparation of polyclonal antibodies].
OBJECTIVE
To express the recombinant proteins of human fibrogenic factors and prepare the polyclonal antibodies of rabbit-anti-human fibrogenic factors.


METHODS
The human fibrogenic factors were variant splicing forms of one gene. Their consensus sequence (ChS) was cloned into plasmid pQE30 to construct recombinant prokaryotic expression system. The recombinant expression vectors were transformed into E.coli M15, and then the expression vectors were induced by IPTG and many insoluble inclusion body proteins was obtained. The obtained proteins were used to immunize rabbits to obtain polyclonal antiserum with high titer and specificity. The eukaryotic expression of the recombinants of human fibrogenic factors was detected by Western blot.


RESULTS
The insoluble inclusion body proteins were also used to prepare polyclonal antibodies with high titer and specificity.


CONCLUSIONS
The expression of the recombinant proteins of human fibrogenic factors and the preparation of polyclonal antiserum provide a useful tool for further study of the expression and function of human fibrogenic factors.
Chen
Xiao-Hua
XH
Department of Biological Science and Biotechnology, Tsinghua University, Beijing, China. cxh01@mails.tsinghua.edu.cn
Cai
Guo-Ping
GP
D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007106:Immune Sera / Q000032:analysis / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D012995:Solubility
D007106:Immune Sera; D011994:Recombinant Proteins