The Preparation and Application of Mouse DEAF1 Polyclonal Antibody

Abstract

Abstract: To study endogenous expression of DEAF1 in mouse pancreatic lymph nodes and the mechanism of DEAF1 regulated expression of peripheral tissue antigen genes. The prokaryotic expression plasmid contain mouse DEAF1 cDNA was transformed into E. coli BL21, the induced recombinant DEAF1 protein was purified by urea gradient cleavage, and was used to immunize Balb/C mouse. The anti-DEAF1 polyclonal antibody can get from mouse serum, and the specificity, sensibility, and affinity of antibody were test by ELISA, Western blot, Immunofluorescence and Immunoprecipitation. The results showed that anti-sera antibody can reacted with antigen and detected a specific band of 60 kD, and also can be used for immunofluorescence localization. Immunoprecipitation experiments showed that anti-sera antibody can bind with the antigen with high affinity. These results suggested that the DEAF1 polyclonal antiserum revealed high specificity, sensibility and affinity.

Key words: Prokaryotic expression and Purification, DEAF1, Animal immunization, Anti-serum Antibody identification

Zhu Xiaohui, Sheng Deqiao, Kang Le, Wu Xianfeng, Shao Wen, Liu Xiaoyan. The Preparation and Application of Mouse DEAF1 Polyclonal Antibody[J]. Biotechnology Bulletin, 2014, 0(5): 197-201.

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