Liquid chemiluminescent DNA pull-down assay to measure nuclear receptor-DNA binding in solution
Liquid chemiluminescent DNA pull-down assay to measure nuclear receptor-DNA binding in solution
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DOI：
10.2144/000112915
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发表时间：
2008-10-01
期刊：
影响因子：
2.7
通讯作者：
Koibuchi, Noriyuki
中科院分区：
文献类型：
--
作者：
Iwasaki, Toshiharu;Miyazaki, Wataru;Koibuchi, Noriyuki
电泳迁移率杆测定（EMSA）通常用于研究体外蛋白质-DNA结合。然而，EMSA可产生相当大量的不期望的废物，特别是当检测有毒化合物时。因此，我们开发了一种新的体外蛋白质-DNA结合试验，称为液体荧光DNA下拉试验，这是基于地高辛标记的DNA和谷胱甘肽融合的DNA结合蛋白结合谷胱甘肽琼脂糖珠之间的溶液杂交。S-转移酶（GST）融合DNA结合蛋白结合谷胱甘肽-琼脂糖珠。
Electrophoretic mobility shaft assays (EMSAs) art commonly used to investigate protein-DNA binding in vitro. However EMSA can generate considerable amounts of undesirable waste, particularly when toxic compounds art, examined. We therefore developed a novel in vitro protein-DNA binding assay called liquid chemiluminescent DNA pull-down assay, which is based on solution hybridization between digoxigenin-labeled DNA and glutathione fused DNA binding protein bound to glutathione-Sepharose beads. S-tranferase (GST)-fused DNA binding protein bound to glutathione-Sepharose beads.