Burke B, Tooze J, Warren G
European Molecular Biology Laboratory, Heidelberg, FRG.
EMBO J. 1983;2(3):361-7. doi: 10.1002/j.1460-2075.1983.tb01431.x.
A monoclonal IgM has been characterised which recognises the nuclear lamins in all mammalian cells tested. In immunoblotting experiments using both one- and two-dimensional gels it recognises lamins A, B and C. The common antigenic determinant lies on a proteolytic fragment of 46,000 daltons which can be generated from each lamin polypeptide by treatment with chymotrypsin. In immunofluorescence experiments on whole cells and thin frozen sections, the antibody labelled only the nuclear envelope and not the nuclear interior. During mitosis, labelling was found dispersed throughout the cell cytoplasm. By immunoelectron microscopy using the antibody and protein A-gold, only the nucleoplasmic side of the nuclear envelope (the nuclear lamina) was labelled, but there was no labelling of the nuclear pores.
已鉴定出一种单克隆IgM,它能识别所有测试的哺乳动物细胞中的核纤层蛋白。在使用一维及二维凝胶的免疫印迹实验中,它能识别核纤层蛋白A、B和C。共同的抗原决定簇位于一个46000道尔顿的蛋白水解片段上,该片段可通过用胰凝乳蛋白酶处理从每个核纤层蛋白多肽产生。在对完整细胞和薄冰冻切片进行的免疫荧光实验中,该抗体仅标记核膜,而不标记核内部。在有丝分裂期间,发现标记物分散在整个细胞质中。通过使用该抗体和蛋白A-金进行免疫电子显微镜观察,仅核膜的核质侧(核纤层)被标记,但核孔没有标记。