Chen Jianfei,Jin Xianqing,Ding Xionghui,Sun Yanhui,Zhao Lihua,Guo Zhenhua,Wang Shiqi.Preparation and identification of polyclonal antibodies of novel multidrug resistance gene HA117[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(11):1128-1131.
Chen Jianfei,Jin Xianqing,Ding Xionghui,Sun Yanhui,Zhao Lihua,Guo Zhenhua,Wang Shiqi.Preparation and identification of polyclonal antibodies of novel multidrug resistance gene HA117[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(11):1128-1131.DOI:
Preparation and identification of polyclonal antibodies of novel multidrug resistance gene HA117
Objective To prepare polyclonal antibodies of a novel multidrug resistance gene
HA117. Methods pAdTrack-HA117 plasmid was used as the template to amplify the total cDNA of HA117 by using PCR. The products of PCR were transferred into BL21 (DE3) competent cells to induce the protein expression of HA117 and identified by SDS-PAGE. New Zealand rabbits were immunized with HA117 protein after been purified in order to get the polyclonal antibodies of HA117. Indirect ELISA was used to detect the titer of the polyclonal antibodies of HA117. Adenovirus vector encoding HA117 gene was transducted into HEK293. Immunofluorescence staining and Western blot analysis were used to detect the specificity of polyclonal antibodies. Results Polyclonal antibodies of HA117 were obtained successfully. The anti-HA117 serum titer was 1∶729 000 by indirect ELISA. The polyclone antibody of HA117 identified the HA117 protein in transducted HEK293 specifically by immunofluorescence staining and Western bolt analysis. Conclusion Our prepared polyclonal antibodies of HA117 are of high titer and high specificity.