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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >SIMULTANEOUS ANALYSIS OF CYCLIN AND ONCOGENE EXPRESSION USING MULTIPLE MONOCLONAL ANTIBODY IMMUNOBLOTS
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SIMULTANEOUS ANALYSIS OF CYCLIN AND ONCOGENE EXPRESSION USING MULTIPLE MONOCLONAL ANTIBODY IMMUNOBLOTS

机译：利用多个单克隆抗体免疫印迹同时分析细胞周期和癌基因的表达

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摘要

Cell dysfunction or dysregulation in cancer generally results from complex gene interactions, numerous cellular events and environmental influences which modify gene expression or post-translational protein modifications. Genetic analysis in itself cannot always predict: or diagnose multigenic diseases. The major technical difficulty is thus to defect, identify and measure simultaneously the expression of several genes and the post-translational modifications of their products. In order to progress io this direction, this paper describes a simple immunoblot method using several monoclonal antibodies to simultaneously analyze oncogene expression and cell cycle specific checkpoints in patient solid biopsies and transformed cell lines. One mg of normal human liver biopsy and HEPG2 (hepatoblastoma-derived cell line) protein samples have been separated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and transferred onto polyvinylidene difluoride (PVDF) membranes. The membranes were stained with amido black, scanned and tested separately with the nine monoclonal antibodies p53, c-myc, PCNA, MEK1, pan-ras, Cip1, Cdc2, Kip1, and TCTP. The nine antibodies of interest were then combined to form a mixture, and simultaneously used as the primary antibobies. This antibody mixture simultaneously detected the nine proteins of interest on both samples and it demonstrated the extensive expression changes and the presence of various isoforms most likely due to post-translational modifications of gene products. [References: 27]
机译：癌症中的细胞功能异常或失调通常是由于复杂的基因相互作用，众多细胞事件和环境影响所致，这些因素改变了基因表达或翻译后蛋白质的修饰。遗传分析本身不能总是预测：或诊断多基因疾病。因此，主要的技术困难是同时缺陷，鉴定和测量几种基因的表达及其产物的翻译后修饰。为了朝这个方向发展，本文描述了一种简单的免疫印迹方法，使用几种单克隆抗体同时分析患者实体活检和转化细胞系中的癌基因表达和细胞周期特异性检查点。一维正常人肝活检和HEPG2（肝母细胞瘤衍生的细胞系）蛋白质样品已通过二维聚丙烯酰胺凝胶电泳（2-D PAGE）分离，并转移到聚偏二氟乙烯（PVDF）膜上。膜用酰胺黑染色，分别扫描并用九种单克隆抗体p53，c-myc，PCNA，MEK1，pan-ras，Cip1，Cdc2，Kip1和TCTP进行测试。然后将九种目的抗体合并以形成混合物，并同时用作一级抗体。该抗体混合物同时在两个样品上检测到9种目的蛋白，并证明了广泛的表达变化和各种同工型的存在，这很可能是由于基因产物的翻译后修饰所致。 [参考：27]

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