[Expression of recombinant human hemangiopoietin and preparation of its polyclonal antibody].
OBJECTIVE
To express the recombinant fusion protein of hemangiopoietin (HAPO) and prepare the rabbit-anti-human HAPO polyclonal antibody.


METHODS
The sequence encoding HAPO was amplified by PCR and cloned into plasmid pET32c to construct recombinant prokaryotic expression system. The recombinant expression vectors were identified by enzyme digestion analysis and transformed into E. coli. The HAPO protein was purified by affinity chromatography. Rabbits were immunized with the HAPO protein, and the immune sera of rabbits were collected. Antibodies (IgG) obtained from the immune sera were purified.


RESULTS
The purified HAPO protein was successfully obtained. The purified polyclonal antibody of rabbit-anti-human HAPO was also obtained from the immune sera of rabbits, and could response to human HAPO.


CONCLUSIONS
A prokaryotic expression system of human HAPO has been prepared and the polyclonal antibody against HAPO has been prepared, which can be used to determine HAPO protein.
Ren
Qian
Q
State Key Laboratory of Experimental Hematology, National Research Center for Stem Cell Engineering and Technology, CAMS and PUMC, Tianjin 300020, China. renqian666@yahoo.com
Liu
Yong-Jun
YJ
Xu
Bin
B
Han
Zhi-Bo
ZB
Lu
Shi-Hong
SH
Ma
Feng-Xia
FX
Chen
Zhong
Z
Han
Zhong-Chao
ZC
D000818:Animals; D015153:Blotting, Western; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera / Q000032:analysis / Q000276:immunology*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011509:Proteoglycans / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000099158:Proteoglycan 4
D004262:DNA Restriction Enzymes; D007106:Immune Sera; D011509:Proteoglycans; D011994:Recombinant Proteins; C419197:PRG4 protein, human; D000099158:Proteoglycan 4