title: [Preparation of polyclonal antibody for heat shock protein 16.3].
abstract: OBJECTIVE
  To induce the prokaryotic expression of His-heat shock protein 16.3 (Hsp16.3) and prepare its polyclonal antibody.
  
  
  METHODS
  DNA primers were designed and synthesized, and Hsp16.3 cDNA was amplified from Mycobacterium tuberculosis H37Rv by PCR. Thereafter, the recombinant vector pET28a-Hsp16.3 was constructed, cloned and induced to express. The His-Hsp16.3 protein was purified using a Ni-NTA column. After identification by SDS-PAGE, the purified protein was used to immunize the New Zealand white rabbits to prepare the polyclonal antibody. The titer and biological activity of the polyclonal antibody were analyzed by ELISA and Western blotting, respectively.
  
  
  RESULTS
  The prokaryotic expression vector pET28a-Hsp16.3 was constructed and expressed well in E.coil BL21(DE3). SDS-PAGE showed that the relative molecular mass was about 20 000. The titer of the polyclonal antibody reached 1:800. The purified antibody was proved to have a good specificity.
  
  
  CONCLUSIONS
  The His-Hsp16.3 has been cloned, expressed and purified successfully. Polyclonal antibody of His-Hsp16.3 protein has been obtained as well.
authors.affiliation: Department of Immunology, Immunology Innovation Base of Postgraduate Education, Zunyi 563003, China.
authors.forename: Shaoying
authors.initials: S
authors.lastname: Gao
authors.affiliation: Department of Immunology, Immunology Innovation Base of Postgraduate Education, Zunyi 563003, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Qin
authors.affiliation: Department of Immunology, Immunology Innovation Base of Postgraduate Education, Zunyi 563003, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Immunology, Immunology Innovation Base of Postgraduate Education, Zunyi 563003, China.
authors.forename: Nalin
authors.initials: N
authors.lastname: Qin
authors.affiliation: Department of Immunology, Immunology Innovation Base of Postgraduate Education, Zunyi 563003, China.
authors.forename: Weina
authors.initials: W
authors.lastname: Yu
authors.affiliation: Department of Immunology, Immunology Innovation Base of Postgraduate Education, Zunyi 563003, China.
authors.forename: Chenbo
authors.initials: C
authors.lastname: Ding
authors.affiliation: Grade 2011 of Undergraduate, Department of Medical Laboratory, Zunyi Medical College, Zunyi 563003, China.
authors.forename: Jianbo
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Grade 2011 of Undergraduate, Department of Nursing, Zunyi Medical College, Zunyi 563003, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Song
authors.affiliation: Department of Immunology, Immunology Innovation Base of Postgraduate Education, Zunyi 563003, China.
authors.forename: Junmin
authors.initials: J
authors.lastname: Luo
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D018833:Chaperonins / Q000235:genetics / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007114:Immunization; D008970:Molecular Weight; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; C431133:Hsp16.3 protein, Mycobacterium tuberculosis; D018833:Chaperonins