抗溴化脱氧尿嘧啶核苷单克隆抗体免疫荧光细胞化学染色过程的最佳条件实验研究  

THE OPTIMAL CONDITION OF THE STAINING PROCEDURE USING MONOCLONAL ANTIBODY TO BROMODEOXYURIDINE (BRDURD)

作　　者：石学耕[1] 西谷巖 善積昇 

机构地区：[1]上海第二医科大学医用激光教研室,上海200025 [2]日本岩手医科大学妇产科

出　　处：《解剖学报》1990年第1期97-101,共5页Acta Anatomica Sinica

摘　　要：抗溴化脱氧尿嘧啶核苷(BrdUrd)单克隆抗体荧光染色过程涉及到以下一些重要步骤:BrdUrd脉冲标记时间长短,核DNA部分变性处理所用盐酸的浓度与处理时间等。应以BrdUrd阳性细胞与阴性细胞之间平均荧光强度之比值,酸处理过程中细胞凝集的比例,细胞群体G_1峰DNA荧光强度及变异系数作为整个染色过程最佳条件的评价标准。在30分钟BrdUrd标记时间,2.4mol/L盐酸30分钟处理时间及抗BrdUrd单克隆抗体20℃1小时温育时间等条件下,可获得满意的染色结果,并且不需要核糖核酸酶的处理。本实验室将KF-1、KFr、HeLa、IK-90等几种培养细胞系用此方法染色均可获得满意的染色结果。The staining procedure using the monoclonal antibody to BrdUrd involved the following important steps: the duration of BrdUrd labeling time, the concentration of HC1, the duration of HCI treatment for cellular DNA partial denaturation and so on. The ratio of mean intensities between BrdUrd positive cells and BrdUrd negative cells, the proportion of cell aggregation occuring during HCI treatment, the relative fluorescence intensity and coefficient of variation (CV) of G_1 peak were considered as the criteria of optimal conditions of the whole staining procedure. The optimal results of this staining procedure were obtained under the conditions of 30 rain BrdUrd labeling time, 2.4 mol/L HC1, 30 min HC1 treatment, 1 hour incubation of the monoclonal antibody to BrdUrd and without RNase treatment. With this staining procedure, the optimal staining results were obtained for KF-1, KFr, HeLa and IK-90 cell lines.

关 键 词：BrdUrd 单克隆抗体 免疫荧光 

分 类 号：R392－33[医药卫生—免疫学]