MONOCLONAL-ANTIBODY TO 5-BROMODEOXYURIDINE AND 5-IODODEOXYURIDINE - A NEW REAGENT FOR DETECTION OF DNA-REPLICATION
MONOCLONAL-ANTIBODY TO 5-BROMODEOXYURIDINE AND 5-IODODEOXYURIDINE - A NEW REAGENT FOR DETECTION OF DNA-REPLICATION
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DOI：
10.1126/science.7123245
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发表时间：
1982-01-01
期刊：
影响因子：
56.9
通讯作者：
GRATZNER, HG
中科院分区：
文献类型：
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作者：
GRATZNER, HG
5-溴脱氧尿苷特异性单克隆抗体已被生产出来并应用于体外逐个细胞检测低水平的 DNA 复制。产生 Ig 的杂交瘤来源于用碘尿苷和卵清蛋白缀合物免疫的小鼠的脾细胞。这些细胞与浆细胞瘤系 SP2/0Ag14 融合。产生的抗体对溴脱氧尿苷和碘脱氧尿苷具有高度特异性，并且不与胸苷发生交叉反应。暴露于溴脱氧尿苷短至 6 分钟的培养细胞中的 DNA 合成可以通过免疫荧光染色方法轻松快速地检测到，并通过流式细胞术进行定量。
Monoclonal antibodies specific for 5-bromodeoxyuridine have been produced and applied in detecting low levels of DNA replication on a cell-by-cell basis in vitro. The Ig-producing hybridomas were derived from spleen cells of mice immunized with a conjugate of iodouridine and ovalbumin. The cells were fused with the plasmacytoma line SP2/0Ag14. The antibodies produced are highly specific for bromodeoxyuridine and iododeoxyuridine and do not cross-react with thymidine. DNA synthesis in cultured cells exposed to bromodeoxyuridine for as short a time as 6 min can be detected easily and rapidly by an immunofluorescent staining method and quantitated by flow cytometry.