Immunoblot specificity of monoclonal antibodies assayed against complex extracts.
Immunoblot specificity of monoclonal antibodies assayed against complex extracts.
复制标题
针对复杂提取物测定单克隆抗体的免疫印迹特异性。
DOI：
10.1016/0003-2697(85)90083-1
复制
发表时间：
1985
影响因子：
2.9
通讯作者：
Fisher,CA
中科院分区：
文献类型：
--
作者：
Lampson,LA;Fisher,CA
使用已知特异性的单克隆抗体和已知抗原表达的靶细胞来评估免疫印迹法对复合物提取物的特异性。抗体组包括10个已知结构抗原的先前表征的单克隆：HLA-A，B，C；Ia;和肌动蛋白。此外，一种商业试剂被证明在免疫印迹中与HLA链反应，而不是与肌动蛋白交叉反应。已知靶细胞有强，弱，或阴性抗原表达进行检查。在所有情况下，结果完全符合抗体、抗原和靶细胞的既定特性。例如，在HLA- a，B，C表达强烈的淋巴样细胞中检测到HLA条带，在淋巴样细胞活性为0.5%的神经细胞系中也检测到HLA条带。没有观察到细胞骨架交叉反应或其他意想不到的特异性条带。这项工作为解释复杂组织中新定义蛋白的单克隆抗体的免疫印迹分析奠定了基础。
Monoclonal antibodies of known specificity, and target cells of known antigen expression, were used to evaluate the specificity of the immunoblot assay against complex extracts. The antibody panel included 10 previously characterized monoclonals to antigens of known structure: HLA-A,B,C; Ia; and actin. In addition, a commercial reagent was shown to react with HLA chains in immunoblots, and not to cross-react with actin. Target cells known to have strong, weak, or negative expression of the antigens were examined. In all cases, the results were in complete accord with the established properties of the antibodies, antigens, and target cells. For example, HLA bands were detected in lymphoid cells with strong HLA-A,B,C expression, and also in a neuronal cell line with 0.5% of the lymphoid activity. Cytoskeletal cross-reactions, or other unexpected specific bands, were not observed. This work lays a foundation for interpreting immunoblot analyses of monoclonal antibodies to newly defined proteins of complex tissue.