棉铃虫类肌钙蛋白多克隆抗体的制备与鉴定

Preparation and Identification of Polyclonal Antibody Against Calponin from Helicoverpa armigera

利用蛋白免疫法制备鼠抗棉铃虫类肌钙蛋白(Helicoverpa armigera Calponin,Ha Cal)的多克隆抗体并进行初步应用。通过原核表达棉铃虫类肌钙蛋白,镍柱纯化获得目的蛋白,经足垫加皮下注射免疫小鼠,4次免疫后用ELISA检测法确定鼠抗Ha Cal蛋白多克隆抗体的效价。通过Western blot和免疫组化检测该多克隆抗体免疫特异性。蛋白免疫法制备的鼠抗Ha Cal多克隆抗体滴度高于1∶819 200。Western-blot和免疫组化显示该抗体能特异性识别棉铃虫体内的天然Ha Cal蛋白。利用蛋白免疫法制备获得了效价高且特异性强的鼠抗棉铃虫Ha Cal多克隆抗体,旨为进一步研究棉铃虫Ha Cal蛋白表达特性及其抗药性功能提供重要的实验工具。

Polyclonal antibodies against calponin from Helicoverpa armigera Ha Cal were prepared by protein immunization and used for preliminary application. The Ha Cal was expressed by the prokaryotic expression system,purified by Ni-column,and then immunized the mice by footpad and subcutaneous injection. The titer of polyclonal antibody against Ha Cal protein was determined by ELISAafter four immunizations. The immunological specificity of the polyclonal antibody was detected by Western blot and immuno histochemical staining. The titer of mouse anti-Ha Cal polyclonal antibody prepared by protein immunization was higher than 1 819 200. Western blot and immuno histochemical staining showed that the antibody specifically recognized the native Ha Cal protein in the H. armigera. In summary,the polyclonal antibody against Ha Cal with high titer and specificity was prepared by protein immunization,which provided an important experimental tool for further studying the expression characteristics of Ha Cal protein and clarifying how to invovle in drug resistance of H. armigera.