Abstract
Co-affinity purification methods can test whether two proteins physically engage in a complex. The assay principle is to enrich cellular extracts for a first protein by a purification step, and then test if a second protein is enriched as well. This principle has been optimized for use at high-throughput in LUMIER (luminescence-based mammalian interactome mapping) assays, which use luciferase-tags for the detection of the second proteins. This protocol is ideal for miniaturization of the assay on microtiter plates and supports a throughput of several hundred interaction tests per week.
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Acknowledgments
We are grateful to Kerstin Mohr and Gabriella Siszler for excellent technical support and to Gabriella Siszler for critical reading of the manuscript and helpful comments.
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Blasche, S., Koegl, M. (2013). Analysis of Protein–Protein Interactions Using LUMIER Assays. In: Bailer, S., Lieber, D. (eds) Virus-Host Interactions. Methods in Molecular Biology, vol 1064. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-601-6_2
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DOI: https://doi.org/10.1007/978-1-62703-601-6_2
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-600-9
Online ISBN: 978-1-62703-601-6
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