Abstract
Sertoli cells are crucial for germ cell support, create a suitable environment for spermatogenesis, and integrate information from the nervous system and germ cell line. To fully understand the role of Sertoli cells, it is necessary to characterize the protein-protein interactions. Identifying the interactome of a given protein may provide leads about the role and molecular mechanism of such protein in Sertoli cells. One of the techniques to characterize protein interactomes consists of co-immunoprecipitation followed by mass spectrometry or Western blot. Co-immunoprecipitation enables the isolation of a protein target and interactome under physiological conditions. In this chapter, we described how to perform an interactomic study using the co-immunoprecipitation technique in Sertoli cells. Moreover, we will focus on how to analyze and interpret the results of a co-immunoprecipitation before mass spectrometry analysis.
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Acknowledgments
This work was supported by FEDER funds through the “Programa Operacional Competitividade e Internacionalização (COMPETE 2020)” and by national funds through the FCT (Fundação para a Ciência e Tecnologia), Projects PTDB/BBB-BQB/3804/2014 and PTDC/DTP-DES/6077/2014. We are thankful to the Institute for Biomedicine (iBiMED) (UID/BIM/04501/2013) for supporting this project. iBiMED is supported by the Portuguese Foundation for Science and Technology (FCT), COMPETE 2020, and FEDER fund. This work was also supported by an individual grant from FCT of the Portuguese Ministry of Science and Higher Education to M.J.F. (SFRH/BD/84876/2012).
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Freitas, M.J., Fardilha, M. (2018). Profiling Signaling Proteins in Sertoli Cells by Co-immunoprecipitation. In: Alves, M., Oliveira, P. (eds) Sertoli Cells. Methods in Molecular Biology, vol 1748. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7698-0_7
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DOI: https://doi.org/10.1007/978-1-4939-7698-0_7
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