title: [Expression and antiserum preparation of recombinant human antithrombin-III].
abstract: OBJECTIVE
  To express recombinant antithrombin-III (AT-III) in an E.coli expression system, prepare the antiserum of AT-III and determine its titer.
  
  
  METHODS
  Gene segment of AT-III was acquired by cloning technology. Then prokaryotic expression plasmid pET32a(+)-AT-III was constructed and transformed into E.coli competent cells BL21. The cells were induced by IPTG to express AT-III. After purified, the product was used to immunize New Zealand rabbits. Then antiserum was detected using indirect ELISA and Western blotting.
  
  
  RESULTS
  Specific bands appeared at about M(r); 77 000, indicating prokaryotic expression protein detected by SDS-PAGE and Western blotting. From the rabbits immunized by the purified fusion protein, we aquired AT-III antiserum, of which the highest titer was 1:12 800 as shown by indirect ELISA. Western blotting showed the antiserum had the ability of specific binding to AT-III protein expressed by 293T and CHO or purified AT-III protein.
  
  
  CONCLUSIONS
  Antiserum of human AT-III has been prepared successfully.
authors: College of Life Science, Northeast Agricultural University, Harbin, China.
authors: Shu
authors: S
authors: Bu
authors: Hongmei
authors: H
authors: Wang
authors: Tao
authors: T
authors: Sun
authors: Wenhao
authors: W
authors: Liu
authors: Liqun
authors: L
authors: Wang
authors: Hongbin
authors: H
authors: He
mesh_terms: D000918:Antibody Specificity; D000990:Antithrombin III / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007106:Immune Sera / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D007106:Immune Sera; D011994:Recombinant Proteins; D000990:Antithrombin III