title: [Preparation and application of polyclonal antibody against mouse IL-1α].
abstract: OBJECTIVE
  To construct a recombinant plasmid encoding mouse IL-1α (mIL-1α), express and purify mIL-1α protein, and prepare its polyclonal antibody.
  
  
  METHODS
  The cDNAs were obtained from the spleen cells of BALB/c mice and the full length of mIL-1α gene was amplified by RT-PCR. Then the mIL-1α gene was inserted into a prokaryotic expression vector pET32a(+) and the resulting recombinant plasmid was transformed into E.coli BL21(DE3). After auto-induction, the mIL-1α protein was expressed and purified by electro-elution. An anti-mIL-1α polyclonal antibody was raised in New Zealand rabbits after immunization with the purified mIL-1α and the titer was determined by ELISA. The specificity of the polyclonal antibody was identified by Western blot and flow cytometry.
  
  
  RESULTS
  The recombinant prokaryotic expression vector pET32a(+)-IL-1α was successfully constructed, and the mIL-1α protein was expressed and purified. ELISA showed the titer of the anti-mIL-1α serum was 1:25 600. Western blot and flow cytometry demonstrated the high specificity of the polyclonal antibody to IL-1α.
  
  
  CONCLUSIONS
  The rabbit anti-mIL-1α polyclonal antibody with high titer and specificity has been prepared after immunization with the purified mIL-1α protein, facilitating further functional studies of IL-1α.
authors: Laboratory of Cellular and Molecular Tumor Immunology, Medical College of Soochow University, Suzhou 215123, China.
authors: Dan-Dan
authors: DD
authors: Lin
authors: Chun-Liang
authors: CL
authors: Liu
authors: Guang-Ming
authors: GM
authors: Bao
authors: Kang
authors: K
authors: Wu
authors: Lin
authors: L
authors: Shan
authors: Hai-Yan
authors: HY
authors: Liu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity / Q000276:immunology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D053582:Interleukin-1alpha / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000906:Antibodies; D053582:Interleukin-1alpha; D011994:Recombinant Proteins