title: [Preparation and characterization of anti-Drosophila Dnop5 antibody].
abstract: OBJECTIVE
  To prepare the rabbit antibody against Dnop5 and identify its specificity.
  
  
  METHODS
  Dnop5 cDNA was amplified by RT-PCR, and then was subcloned into the fusion expression vectors pET28a(+). After being expressed in E.coli BL21, the truncated Dnop5 protein was purified and used to immunize rabbit. Purified antibody was obtained through affinity chromatography column with the expressed Dnop5. The specificity of the purified antibody was characterized by Western blot and immunohistochemical staining.
  
  
  RESULTS
  The Dnop5 gene was successfully inserted into pET28a(+). After induction, the fusion protein was expressed in the form of inclusion body. The purified fusion protein was obtained by affinity chromatography. After immunization of rabbits, the antibody against Dnop5 was obtained. Western blot analysis and immunohistochemical staining showed that the antibody had a good specificity.
  
  
  CONCLUSIONS
  The rabbit antibody against Dnop5 has been successfully prepared, which lays the foundation for further study on the Dnop5 function.
authors.affiliation: School of Basic Medical Science, Key Laboratory of Developmental Genes and Human Diseases, Ministry of Education, Southeast University, Nanjing 210009, China. yld@seu.edu.cn
authors.forename: Liu-di
authors.initials: LD
authors.lastname: Yuan
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yong-qi
authors.initials: YQ
authors.lastname: Wan
authors.forename: Jie
authors.initials: J
authors.lastname: Ding
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology*; D000918:Antibody Specificity; D004330:Drosophila; D029721:Drosophila Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D020718:Ribonucleoproteins, Small Nucleolar / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000906:Antibodies; D029721:Drosophila Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D020718:Ribonucleoproteins, Small Nucleolar; C499061:nop5 protein, Drosophila