Assessment of small ligand-protein interactions by electrophoretic mobility shift assay using DNA-modified ligand as a sensing probe
Assessment of small ligand-protein interactions by electrophoretic mobility shift assay using DNA-modified ligand as a sensing probe
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DOI：
10.1007/s10529-006-9301-6
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发表时间：
2007-05-01
影响因子：
2.7
通讯作者：
Kobatake, Eiry
中科院分区：
文献类型：
--
作者：
Funabashi, Hisakage;Ubukata, Michito;Kobatake, Eiry
通过电泳迁移率变动分析来评估小配体与蛋白质之间的相互作用。通过用DNA修饰模型配体生物素来产生传感探针。DNA修饰配体与抗生物素抗体或链霉亲和素作为靶蛋白的复合物通过琼脂糖凝胶电泳进行分析。在靶蛋白存在下，对应于DNA修饰的配体的条带发生位移，通过加入浓度从0.1 μ M到100 μ M的游离配体，位移条带的强度降低。从该校准可以确定样品中配体的浓度，从而允许评估小配体与其靶之间的相互作用。
The interaction between a small ligand and a protein were assessed by electrophoretic mobility shift assay. A sensing probe was created by modifying the model ligand, biotin, with DNA. The complex of DNA-modified ligand and anti-biotin antibody or streptavidin as a target protein was analyzed by agarose gel electrophoresis. The band corresponding to the DNA-modified ligand was shifted in the presence of the target protein, and the intensities of the shifted bands were decreased by adding increasing concentrations of free ligand ranging from 0.1 mu M to 100 mu M. From this calibration the concentration of ligand in the samples could be determined, allowing for evaluation of the interaction between a small ligand and its target.