title: [Preparation of anti-BRDT-NY polyclonal antibody and expression of BRDT-NY protein in digestive tract tumors].
abstract: OBJECTIVE
  To prepare the mouse polyclonal antibody against human BRDT-NY prokaryotic protein and analyze the expression of BRDT-NY protein in digestive tract tumors.
  
  
  METHODS
  The N-terminal amino acids of BRDT-NY protein was amplified by PCR and cloned into the expression vector pET28a(+). The recombinant plasmid pET28a+-BRDT-NY was transformed into E.coli BL21 and induced to express the recombinant protein with IPTG. We immunized BALB/c mice with the purified BRDT-NY protein for preparing the specific polyclonal antibody. The titer of the antibody was analyzed by ELISA. The expression of BRDT-NY protein in digestive tract tumors was detected by immunohistochemistry.
  
  
  RESULTS
  Recombinant human BRDT-NY protein was expressed in BL21 and purified successfully by Ni-affinity chromatography. Two months after the mice were immunized with the purified BRDT-NY protein, we obtained the specific polyclonal antibody of high titer 1:100 000. Immunohistochemical analysis revealed that BRDT-NY protein was highly expressed in digestive tract tumors.
  
  
  CONCLUSIONS
  The successful preparation of mouse polyclonal antibody against BRDT-NY lays a foundation for the research on the role of BRDT-NY protein in the pathology of human digestive tract tumors.
authors.affiliation: Medical Centre, Medical College, Yangzhou University, Yangzhou, China. wmding@yzu.edu.cn
authors.forename: Wei-min
authors.initials: WM
authors.lastname: Ding
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zheng
authors.forename: Hong-ya
authors.initials: HY
authors.lastname: Sun
authors.forename: Hong
authors.initials: H
authors.lastname: Liang
authors.forename: Jian-jun
authors.initials: JJ
authors.lastname: Wang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D005770:Gastrointestinal Neoplasms / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009687:Nuclear Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D000092468:Bromodomain Containing Proteins
chemical_list: D000906:Antibodies; D009687:Nuclear Proteins; C109570:BRDT protein, human; D000092468:Bromodomain Containing Proteins