Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (12): 141-146.doi: 10.13560/j.cnki.biotech.bull.1985.2014.12.023

• Research Report • Previous Articles     Next Articles

Preparation of the Polyclonal Antibody Against Helicoverpa armigera FK506 Binding Protein by Immunizing with pcDNA3-FK506BP

Zhu Yan, Zhang Fuchun, Ma Ji, Huang Li’na, Liu Xiaoning   

  1. (Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046)
  • Received:2014-03-31 Online:2014-12-08 Published:2014-12-12

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Abstract: It was to prepare the polyclonal antibody against FK506 binding protein, which was to be to verified the specificity of the antibody by Western blot. Firstly, recombinant expression plasmid pcDNA3-FKBP12 was constructed and sequenced, and the recombinant plasmid pcDNA3-FKBP12 was then transfered into Kunming mice four times. Finally, ELISA assay, Western blot analysis and immunohistochemistry(IHC)were used to determine the titer of polyclonal antibody of FK506 binding protein and its specificity, respectively. The results showed that the titer of polyclonal antibody reached 1∶25 600, and it was able to bind specifically to both the fusion protein His- FKBP12 and FKBP12 from the different tissues of the cotton bollworm in the 3rd larvae, which laid the foundation for the subsequent experiments.

Key words: Helicoverpa armigera, FK506 binding protein, Polyclonal antibody