小鼠抗重组山羊胎盘催乳素多克隆抗体的制备及其在山羊胎盘组织中定位的研究  

Preparation of anti-recombant caprine placental lactogen polyclonal antibody and its localization in caprine placenta tissue

作　　者：梁庆龙[1] 浩洪龙[1] 牟瑞营[1] 王林峰[1] 葛利江[1] 

机构地区：[1]山东农业大学动物科技学院临床兽医系,山东泰安271018

出　　处：《西南农业学报》2008年第1期183-186,共4页Southwest China Journal of Agricultural Sciences

摘　　要：用rcPL对6-8周龄的昆明小鼠进行免疫注射制备多抗，第3次加强免疫后第6天采集少许血清，用琼脂凝胶免疫扩散（AGID）和间接酶联免疫吸附试验（ELISA）检测抗体的产生情况；分娩后立即通过剖腹产的方法获得山羊胎盘组织，用获得的抗血清通过免疫组织化学（IHC）和间接荧光抗体试验（IFA）的方法定位胎盘催乳素。结果表明，琼脂扩散检测抗原浓度为100μg／mL，用ELISA中酶标仪检测450nm处OD值，免疫血清与阴性对照的比值P／N为7．77；IHC和IFA显示胎盘催乳素在肉阜和子叶的双核细胞及单核细胞中表达。使用背部皮下多点注射的方法对昆明小白鼠进行免疫来制备重组山羊胎盘催乳素多克隆抗体是可行的，制备的抗体能满足免疫组织化学和荧光免疫组织化学检测胎盘催乳素的要求。Polyclonal antibody was prepared in 6 to 8 week-old Kunming mouse. Serum was collected on the sixth days after third boost immunization, and the concentration of antiserum was detected with agar gel immunodiffusion（AGID） and indirect enzyme-linked immunosorbent assay（ELISA） ; Placental tissue were obtained after postpartum by caesarean section and was localized of placental lactogen by the methods of Immunohistochemistry and indirect fluorescence antibody assay （IFA）. The antigen could be detected at a concentration of 100 μg/mL by the method of AGID, and the P/N data was 7.77 by the method of ELISA ; It was obvious that the caprine placental lactogen was expressed in the binucleate and uninucleate cells of the carunclcs and cotyledons by immunohistochemical and immunofluorecence methods. It was possible to raise polyclonal antibody of rcPL by the Kunmiug mouse in the way of injecting subcutaneously in multiple sites along theback,and the preparative antibody could be used to examine PL in placenta by the methods of IHC and IFA.

关 键 词：rcPL 山羊胎盘组织 多克隆抗体 AGID ELISA IHC IFA 

分 类 号：S852.4[农业科学—基础兽医学]