Validation of Isoform- and Mutation-Specific RAS Antibodies.
Validation of Isoform- and Mutation-Specific RAS Antibodies.
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同种型和突变特异性 RAS 抗体的验证。
DOI：
10.1007/978-1-0716-1190-6_5
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Der,ChanningJ
中科院分区：
文献类型：
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作者：
Waters,AndrewM;Der,ChanningJ
抗体特异性的验证对于准确评估蛋白表达至关重要。对于识别RAS癌基因家族基因产物（HRAS、KRAS和NRAS）的抗体来说，一个重要的挑战是确定四种几乎相同的HRAS、KRAS4A、KRAS4B和NRAS蛋白的选择性。随着人们对不同RAS蛋白在正常细胞和肿瘤细胞中的不同作用的认识不断增加，需要经过良好验证的抗体来评估不同RAS亚型的功能和表达。在这里，我们描述了我们的实验方法，以表征RAS抗体的异构体和突变特异性，用于免疫印迹分析。
Validation of antibody specificity is essential for the accurate evaluation of protein expression. For antibodies that recognize the gene products of the RAS family of oncogenes (HRAS,KRAS,andNRAS), an important challenge is the determination of selectivity for the four nearly identical HRAS, KRAS4A, KRAS4B, and NRAS proteins. With increasing appreciation for the distinct roles of the different RAS proteins in normal and neoplastic cells, there is a need for well-validated antibodies to evaluate the function and expression of the different RAS isoforms. Here we describe our experimental approaches to characterize RAS antibodies for their isoform- and mutant-specificity for use in immunoblot analyses.