Abstract
Co-immunoprecipitation is an invaluable technique in evaluating native protein–protein interactions in vitro and in vivo. However, it can be difficult to detect interactions of a very transient nature, particularly interactions with phosphatases and kinases. The evaluation of the phosphorylation status of c-Myc can also be challenging with the current commercially available phosphorylation sensitive antibodies. Here, we describe two protocols: one for the co-immunoprecipitation of endogenous c-Myc to detect protein–protein interactions and second, for the immunoprecipitation of endogenous c-Myc to probe for phosphorylation status.
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Acknowledgments
The authors would like to thank Julienne R. Escamilla-Powers, Deanne Tibbits, and Hugh K. Arnold for a helpful discussion. This work was supported by Department of Defense Breast Cancer Research Program Award BC061306, Susan G. Komen Breast Cancer Foundation Awards BCTR0201697 and BCTR0706821, and National Institutes of Health Awards 1 R01 CA129040 and CA100855 to R.C.S.
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Daniel, C.J., Zhang, X., Sears, R.C. (2013). Detection of c-Myc Protein–Protein Interactions and Phosphorylation Status by Immunoprecipitation. In: Soucek, L., Sodir, N. (eds) The Myc Gene. Methods in Molecular Biology, vol 1012. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-429-6_5
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DOI: https://doi.org/10.1007/978-1-62703-429-6_5
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