Sertoli Cell Preparation for Co-immunoprecipitation
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Most techniques to study protein-protein interactions, gene expression or signal transduction, among others, in Sertoli cells, depend on obtaining a protein extract of such cells. This is accomplished by lysing the Sertoli cells and solubilizing the intracellular proteins. Depending on the purpose of your study, the technique used to lyse and consequently obtain protein extracts from Sertoli cells must be considered. In this chapter, we will focus on how to obtain a protein Sertoli cell extract suitable for co-immunoprecipitation technique.
KeywordsLysis buffer Protein quantification Co-immunoprecipitation Sertoli cells Western blot
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), Compete2020, 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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