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1. 上海第二医科大学
2. 无锡核医学国家重点实验室
3. 中国科学院上海细胞生物学研究所
4. 上海第二医科大学无锡核医学国家重点实验室中国科学院上海细胞生物学研究所
纸质出版日期:1995-05-01
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[1]陆敏,照日格图,何荣霞,易宁育,夏宗勤.M受体亚型放射配基结合-免疫沉淀分析方法的建立及考核[J].核技术,1995(05):263-267.
Establishment of a radioligand binding-immunoprecipitation technique for the assay of subtypes of M-cholinergic receptors[J]. Nuclear techniques, 1995, (5).
利用1%洋地黄皂甙溶解大鼠脑匀浆M受体,用非选择性配基3H-QNB标记可溶性M受体,以自制的M1、M2亚型抗血清和固相二抗分别特异性地免疫沉淀相应的亚型,分析单一受体亚型。对方法进行了考核,包括受体的溶解效率、抗原抗体反应时间、抗血清稀释度、反应温度等对分析结果的影响。对大鼠脑M受体的试验表明,用两种亚型抗血清进行的免疫沉淀分析呈典型的单位点饱和曲线,抗体对配基与受体的结合反应无影响。
Specific polyclonal antibodies against M1 and M2 subtypes of cholinergic receptors were prepared in this laboratory.The M-cholinoceptor molecules in brain homogenate were solubilized with 1% digitonin and labelled with 3H-QNB prior to immumuprecipitation. One of the specific antibodies was then added to form antigen-antibody complex with the respective subtype and the complex was precipitated by the aid of a second antibody. Thus the receptor-ligand binding parameters of a single subtype were obtained from the radioactivities of the precipitates.Typical single- site saturation curves were obtained for both M1 and M2 subtypes.No influence of the antibodies on receptor-ligand binding was detected.Results of studies on the solubilization of menbrane receptors
the time course of antibody-antigen reaction
the optimal dilution factor and the reaction temperature are also presented in the paper.
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