Detecting p53 isoforms at protein level.
Detecting p53 isoforms at protein level.
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DOI：
10.1007/978-1-62703-236-0_2
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发表时间：
2013-01-01
期刊：
影响因子：
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通讯作者：
Bourdon, Jean-Christophe
中科院分区：
文献类型：
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作者：
Marcel, Virginie;Khoury, Marie P;Bourdon, Jean-Christophe
人 p53 蛋白亚型在多种细胞系中表达，并主要通过调节基因表达来调节 p53 肿瘤抑制活性 (1-4)。因此，识别细胞系中 p53 同工型的表达模式是 p53 网络未来研究的关键步骤 (5)。目前，p53 蛋白亚型的检测基于一组抗体的使用，通过比较它们的分子量和不同抗体的检测模式来识别它们 (6)。在此，描述了补充有技术知识的经典方案，通过蛋白质印迹和免疫沉淀在蛋白质水平上检测 p53 蛋白亚型。此外，还提供了一种通过荧光素酶报告基因测定来研究 p53 蛋白亚型对 p53 转录活性影响的简单方法。
The human p53 protein isoforms are expressed in several cell lines and modulate p53 tumor suppressor -activity, mainly through modulation of gene expression (1-4). Thus, identifying the pattern of p53 isoforms expression in cell lines is a key step for future studies of the p53 network (5). At the moment, the detection of p53 protein isoforms is based on the use of a panel of antibodies allowing their identification by comparing their molecular weights and their detection pattern by different antibodies (6). Here, classical protocols supplemented with technical know-how are described to detect p53 protein isoforms at protein level by Western blotting and immunoprecipitation. Furthermore, a simple method to study the impact of p53 protein isoforms on p53 transcriptional activity through luciferase reporter gene assays is provided.