Analysis of O6-methylguanine-DNA methyltransferase in individual human cells by quantitative immunofluorescence microscopy.
Analysis of O6-methylguanine-DNA methyltransferase in individual human cells by quantitative immunofluorescence microscopy.
复制标题
通过定量免疫荧光显微镜分析单个人类细胞中的 O6-甲基鸟嘌呤-DNA 甲基转移酶。
作者：
M. Belanich;T. Ayi;B. F. Li;J. Kibitel;D. Grob;T. Randall;A. White;M. Citron;D. Yarosh
描述了一种免疫荧光定量测定，可以使用荧光显微镜对来自标准病理标本的单个细胞进行。该技术已应用于测量 O6-甲基鸟嘌呤-DNA 甲基转移酶，这是一种 DNA 修复蛋白，是癌症化疗中使用的氯乙基硝基源耐药性的分子标记。免疫荧光测定利用对人转移酶具有特异性的单克隆抗体、具有数字成像的荧光显微镜、荧光珠内标和计算机图像分析。该方法专门针对转移酶，产生与活性测量相关的结果，并产生有关组织异质性和亚细胞定位的新数据，而这些数据以前无法通过标准测定方法获得。
A quantitative assay of immunofluorescence is described that can be performed on individual cells from standard pathologic specimens using fluorescence microscopy. The technique has been applied to measurement of O6-methylguanine-DNA methyltransferase, a DNA repair protein that is a molecular marker for resistance to chloroethylnitrosources used in cancer chemotherapy. The immunofluorescence assay makes use of monoclonal antibodies with specificity for human transferase, fluorescence microscopy with digital imaging, fluorescent bead internal standards, and computerized image analysis. This method is specific for the transferase, produces results correlated with activity measurements, and yields new data about tissue heterogeneity and subcellular localization previously unavailable with standard assay methods.