Wu Y H, Franden M A, Hawker J R, McHenry C S
J Biol Chem. 1984 Oct 10;259(19):12117-22.
We have established three murine hybridoma cell lines that secrete monoclonal antibodies directed specifically against the alpha subunit of the DNA polymerase III holoenzyme of Escherichia coli. All three antibodies have been purified and identified to be of the IgM class. competition binding assays indicate that these antibodies bind to at least two distinct but adjacent or interacting sites. An immunoblot assay has been developed that permits quantitation of alpha in crude extracts. These assays agree with previous determinations of the number of polymerase molecules present/cell and indicate that the size of alpha is the same in both the crude and pure form. Anti-alpha IgM can be used with fixed Staphylococcus aureus, cells and anti-IgM to specifically precipitate alpha.
我们已经建立了三种小鼠杂交瘤细胞系，它们分泌的单克隆抗体能特异性地针对大肠杆菌DNA聚合酶III全酶的α亚基。所有这三种抗体均已纯化，并鉴定为IgM类。竞争结合试验表明，这些抗体至少结合到两个不同但相邻或相互作用的位点。已经开发出一种免疫印迹试验，可对粗提物中的α进行定量。这些试验与先前对每个细胞中存在的聚合酶分子数量的测定结果一致，并表明α在粗提物和纯品中的大小相同。抗α IgM可与固定的金黄色葡萄球菌、细胞和抗IgM一起使用，以特异性沉淀α。
