Abstract
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.
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Acknowledgments
This work was supported by NIH grants R01 CA241661 (Tsourkas) and R44 EB023750 (Yu). Andrew Tsourkas is a founder and owns equity in AlphaThera, a biotechnology company that sells antibody conjugation products.
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Niu, J., Hagen, J., Yu, F., Kalyuzhny, A.E., Tsourkas, A. (2023). Labeling of Phospho-Specific Antibodies with oYo-Link® Epitope Tags for Multiplex Immunostaining. In: Kalyuzhny, A.E. (eds) Signal Transduction Immunohistochemistry. Methods in Molecular Biology, vol 2593. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2811-9_7
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DOI: https://doi.org/10.1007/978-1-0716-2811-9_7
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