Kilmartin J V, Wright B, Milstein C
J Cell Biol. 1982 Jun;93(3):576-82. doi: 10.1083/jcb.93.3.576.
Hybrid myeloma cell lines secreting monoclonal antibodies to tubulin have been prepared using rat myelomas and spleen cells from rats immunized with yeast tubulin. A comparison between the results obtained with the rat myeloma Y3-Ag 1.2.3., which secretes a light chain, and a new line, YB2/O, which does not, shows that they are both excellent parental lines and that the second produces hybrids with no myeloma chain components. The antitubulin antibodies in the serum of rats bearing two of the hybrid myeloma tumors gave titers of up to 1:10(6) from which large amounts of monoclonal antibodies could be easily purified. They recognized tubulin from yeast as well as from birds and mammals. The two antibodies gave clear immunofluorescent staining of yeast mitotic spindles as well as the interphase microtubule network of tissue culture cells. Some difference in the pattern of immunofluorescence staining of yeast cells and nuclei was observed between the two antibodies. The purified antibodies could be conjugated to colloidal gold particles and used for direct labeling of yeast microtubules for electron microscopy.
利用大鼠骨髓瘤细胞和用酵母微管蛋白免疫的大鼠脾脏细胞，制备了分泌抗微管蛋白单克隆抗体的杂交骨髓瘤细胞系。对分泌轻链的大鼠骨髓瘤细胞Y3-Ag 1.2.3.和不分泌轻链的新细胞系YB2/O的实验结果进行比较，发现它们都是优良的亲代细胞系，且后者产生的杂交细胞没有骨髓瘤链成分。携带两种杂交骨髓瘤肿瘤的大鼠血清中的抗微管蛋白抗体效价高达1:10(6)，从中可轻松纯化大量单克隆抗体。这些抗体能识别来自酵母以及鸟类和哺乳动物的微管蛋白。这两种抗体对酵母有丝分裂纺锤体以及组织培养细胞的间期微管网络都产生清晰的免疫荧光染色。观察到这两种抗体对酵母细胞和细胞核的免疫荧光染色模式存在一些差异。纯化后的抗体可与胶体金颗粒偶联，用于电子显微镜下酵母微管的直接标记。
