磷酸化
化学
蛋白质磷酸化
信号转导
表位
细胞生物学
荧光蛋白
生物化学
靶蛋白
计算生物学
抗体
蛋白激酶A
生物
绿色荧光蛋白
基因
免疫学
作者
Pan Li,Anton Iliuk,Shuai Yu,Robert L. Geahlen,W. Andy Tao
摘要
We report here for the first time the multiplexed quantitation of phosphorylation and protein expression based on a functionalized soluble nanopolymer. The soluble nanopolymer, pIMAGO, is functionalized with Ti (IV) ions for chelating phosphoproteins in high specificity and with infrared fluorescent tags for direct, multiplexed assays. The nanopolymer allows for direct competition for epitopes on proteins of interest, thus facilitating simultaneous detection of phosphorylation by pIMAGO and total protein amount by protein antibody in the same well of microplates. The new strategy has a great potential to measure cell signaling events by clearly distinguishing actual phosphorylation signals from protein expression changes, thus providing a powerful tool to accurately profile cellular signal transduction in healthy and disease cells. We anticipate broad applications of this new strategy in monitoring cellular signaling pathways and discovering new signaling events.
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