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文献和实验激活转录因子-5(Activating transcription factor-5,ATF5)在黑素瘤中高度表达,其可促进肿瘤细胞的存活。作者利用基因组规模的RNAi筛选方法,鉴定了ATF5的转录调节因子。 结果揭示了黑素瘤细胞存活所必需的一条重要信号传导途径,即RAS-mitogen激活蛋白激酶和磷酸肌醇-3激酶信号盒的激活,可以诱导转录因子CREB3L2(cAMP response element–binding protein-3–like-2),从而直接激活
: 1. Prepare Whole Cell Extract. Make sure every solution DOES NOT contain DTT. 2. Add 1 or 2 μl of non-diluted monoclonal antibody (4B4 is better than 10H8 for HSF1) or 1/10 diluted polyclonal antibody to 10 μg of whole cell extract
Western Analysis of Transgenic Plants
are transferred to a solid support, which is normally nitrocellulose or polyvinylidene fluoride (PVDF [Millipore, Bedford, MA]). Immunological detection of proteins transferred to a solid support combines ease of handling with accessibility of antibodies
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