山羊抗猪脂肪细胞膜蛋白抗体的制备与ELISA鉴定  

Preparation of Antisera in Goat to Pig Adipocyte Membrane Protein and Its Appraisal by Enzyme-linked Immunosorbent Assay

作　　者：赵伟[1] 唐宝定[2] 黄夺先[1] 陈杰[3] 

机构地区：[1]江苏省农业科学院畜牧兽医研究所,南京210014 [2]安徽省蚌埠医学院,蚌埠233003 [3]南京农业大学动物医学院,南京210095

出　　处：《江苏农业学报》2000年第2期97-101,共5页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省自然科学基金资助项目! (BK95 0 5 930 2 )

摘　　要：猪屠宰后 ,迅速采取其皮下脂肪在 37℃下的膜提取液中的匀浆 ,于 37℃温箱中孵育 30 min后 ,用差速离心法和高速离心法提取脂肪细胞膜蛋白 ,并作 SDS- PAGE电泳分析脂肪细胞膜蛋白与其它组织细胞膜蛋白的差异。从电泳图上看出 ,脂肪细胞有很多特异膜蛋白 ,也有一些与其它组织细胞膜蛋白相似的条带。膜蛋白与佐剂充分乳化后主动免疫 3只山羊 ,免疫后 70 d取血清 ,用 EL ISA检测抗体效价为 1:6 40 0。同样 ,用 EL ISA测定抗体与其它组织细胞膜的交叉反应 ,结果显示 :抗猪脂肪细胞膜抗体与其它组织细胞膜有交叉反应 ,但反应性不高。The adipose tissue extracted from crossbred pigs(50～80 kg)was homogenized in 37℃ membrane extractant.After that the homogenate was incubated at 37℃ for 30 minutes.Then the membrane protein of pig adipocyte was normally obtained by centrifugation for several times.Comparative analyses of membrane protein profiles between adipocyte and other tissue cell proteins was conducted by SDS-PAGE.It showed that adipcoyte possessed many specific protein profiles and some ones were similar to other tissue cells.The goat antisera against pig adipocyte membrane protein was produced by multipoint subcutaneous injection of membrane antigen in Freunds adjuvant.Goat antisera were obtained 70 days after the first booster injection and the specific antibodies titer reached 1∶6 400 detected by ELISA.Low cross-reactivity of the antisera to other pig tissues could be detected.

关 键 词：猪脂肪细胞 膜蛋白抗体 免疫 山羊 ELISA鉴定 

分 类 号：S858.272.2[农业科学—临床兽医学] S852.21[农业科学—兽医学]