[Preparation and identification of the rat antibody against recombinant buckwheat trypsin inhibitor]
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Published on: December 14, 2017
Yu-ying Li1, Xiao-dong Cui, Zheng Zhang
1Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China.
Aim:
To prepare the rat antibody against the recombinant buckwheat trypsin inhibitor.
Methods:
The recombinant buckwheat trypsin inhibitor was expressed in E.coli. BL21 (DE3) and used as an immunogen to immunize the rat. The titer and specificity of the anti-BTI antibody from the rat were analyzed by ELISA, Western blot and immunohistochemistry, respectively.
Results:
ELISA detection indicated the titer of the antiserum was about 1:128 000. Western blot analysis showed the antibody reacted specifically with rBTI. The immunohistochemistry analysis proved that rBTI was expressed in the cytoplast and nucleus of EC9706 cells.
Conclusion:
The rat antibody against rBTI has high titer and specificity, which is beneficial to further study on the molecular mechanisms in rBTI-induced apoptosis of tumor cells and also provide an important theory basis for further exploring the relationship between the structure and function of BTI.
