[Preparation and identification of the rat antibody against recombinant buckwheat trypsin inhibitor].
OBJECTIVE
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.


CONCLUSIONS
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.
Li
Yu-ying
YY
Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China.
Cui
Xiao-dong
XD
Zhang
Zheng
Z
Wang
Zhuan-hua
ZH
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D019612:Fagopyrum / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007150:Immunohistochemistry; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
D000906:Antibodies; D007106:Immune Sera; D010940:Plant Proteins; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors