title: [Expression of recombinant human hemangiopoietin and preparation of its polyclonal antibody].
abstract: 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.
authors.affiliation: State Key Laboratory of Experimental Hematology, National Research Center for Stem Cell Engineering and Technology, CAMS and PUMC, Tianjin 300020, China. renqian666@yahoo.com
authors.forename: Qian
authors.initials: Q
authors.lastname: Ren
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Liu
authors.forename: Bin
authors.initials: B
authors.lastname: Xu
authors.forename: Zhi-Bo
authors.initials: ZB
authors.lastname: Han
authors.forename: Shi-Hong
authors.initials: SH
authors.lastname: Lu
authors.forename: Feng-Xia
authors.initials: FX
authors.lastname: Ma
authors.forename: Zhong
authors.initials: Z
authors.lastname: Chen
authors.forename: Zhong-Chao
authors.initials: ZC
authors.lastname: Han
mesh_terms: 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
chemical_list: D004262:DNA Restriction Enzymes; D007106:Immune Sera; D011509:Proteoglycans; D011994:Recombinant Proteins; C419197:PRG4 protein, human; D000099158:Proteoglycan 4