DIABLA: a new screening method for the discovery of protein targets.
DIABLA: a new screening method for the discovery of protein targets.
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DIABLA：一种发现蛋白质靶标的新筛选方法。
DOI：
10.1021/pr800354f
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发表时间：
2008
影响因子：
4.4
通讯作者：
Tolley,Luke
中科院分区：
文献类型：
--
作者：
Montgomery,Robbie;Shay,Hanna;McCarroll,Matthew;Tolley,Luke
动态等电/各向异性结合配体分析（DIABLA）是一种新的方法，用于鉴定复杂样品中与目标分子结合的蛋白质。这是通过首先使用毛细管等电聚焦（cIEF）在毛细管中基于其等电点分离蛋白质来实现的。当待测化合物存在于分离缓冲液中时进行该分离。当蛋白质被聚焦时，整个毛细管被扫描以识别非零各向异性的区域，这些区域是测试化合物与聚焦的蛋白质条带相互作用的位置。通过观察荧光素标记的孕酮与MCF-7乳腺癌细胞裂解物的结合来证明DIABLA。用罗丹明标记蛋白质以允许其观察，然后在标记的孕酮存在下聚焦。各向异性测量表明，孕酮结合到样品中的六个不同的蛋白质带。
Dynamic isoelectric/anisotropy binding ligand assay (DIABLA) is a new method to identify proteins in a complex sample that bind to a molecule of interest. This is accomplished by first using capillary isoelectric focusing (cIEF) to separate the proteins in a capillary based on their isoelectric point. This separation is performed while the compound being tested is present in the separation buffer. When the proteins are focused, the entire capillary is scanned to identify regions of nonzero anisotropy, which are locations where the test compound is interacting with a focused protein band. DIABLA was demonstrated by observing the binding of fluorescein-tagged progesterone to an MCF-7 breast cancer cell lysate. The proteins were tagged with rhodamine to permit their observation and then focused in the presence of the tagged progesterone. Anisotropy measurements show that progesterone binds to six different proteins bands in the sample.