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.