Labeling of Phospho-Specific Antibodies with oYo-Link® Epitope Tags for Multiplex Immunostaining.
Labeling of Phospho-Specific Antibodies with oYo-Link® Epitope Tags for Multiplex Immunostaining.
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使用 oYo-Link® 表位标签标记磷酸化特异性抗体，用于多重免疫染色。
DOI：
10.1007/978-1-0716-2811-9_7
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Tsourkas,Andrew
中科院分区：
文献类型：
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作者：
Niu,Jingwen;Hagen,Jodi;Yu,Feifan;Kalyuzhny,AlexanderE;Tsourkas,Andrew
空间蛋白质组学最近引起了人们的极大兴趣，因为它通过将蛋白质表达模式从亚细胞水平连接到组织甚至生物体水平，为健康和疾病的生物过程提供了前所未有的见解。这些高含量的方法通常依赖于高度的多路复用，从而可以同时检测多种蛋白质。最通用的多路复用方法利用抗体来赋予对各种感兴趣的细胞内蛋白的特异性。因此，这些方法必须能够一次区分许多抗体。在这一章中，我们描述了一个简单而快速的方法来标记抗体与不同的表位标签在一个特定的位点的方式。这允许多个抗体，甚至来自相同的宿主物种，被唯一地识别和检测，并提供了一个简单的方法用于空间蛋白质组学应用。
Spatial proteomics has recently garnered significant interest, as it offers to provide unprecedented insight into biological processes in both health and disease, by connecting protein expression patterns from the subcellular level to the tissue or even organism level. These high-content approaches generally rely on a high degree of multiplexing, whereby multiple proteins can be detected simultaneously. The most versatile multiplexing approaches utilize antibodies to confer specificity for various intracellular proteins of interest. Therefore, these methods must be able to differentiate many antibodies at once. In this chapter, we describe a simple and rapid approach to labeling antibodies with distinct epitope tags in a site-specific manner. This allows multiple antibodies, even from the same host species, to be uniquely identified and detected and offers a simple approach for spatial proteomic applications.