Abstract
The human p53 protein isoforms are expressed in several cell lines and modulate p53 tumor suppressor activity, mainly through modulation of gene expression (1–4). Thus, identifying the pattern of p53 isoforms expression in cell lines is a key step for future studies of the p53 network (5). At the moment, the detection of p53 protein isoforms is based on the use of a panel of antibodies allowing their identification by comparing their molecular weights and their detection pattern by different antibodies (6). Here, classical protocols supplemented with technical know-how are described to detect p53 protein isoforms at protein level by Western blotting and immunoprecipitation. Furthermore, a simple method to study the impact of p53 protein isoforms on p53 transcriptional activity through luciferase reporter gene assays is provided.
Virginie Marcel and Marie P. Khoury have contributed equally to this work.
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Acknowledgments
This work was supported by Cancer Research UK (C8/A6613). V.M. is supported by Breast Cancer Campaign, M.P.K., K.F., A.D., and J.C.B. are supported by Cancer Research UK.
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Marcel, V., Khoury, M.P., Fernandes, K., Diot, A., Lane, D.P., Bourdon, JC. (2013). Detecting p53 Isoforms at Protein Level. In: Deb, S., Deb, S. (eds) p53 Protocols. Methods in Molecular Biology, vol 962. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-236-0_2
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DOI: https://doi.org/10.1007/978-1-62703-236-0_2
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