Abstract
The cell is a fantastic place where molecules dynamically move through the various cellular structures and compartments and meet each other, either transiently or in more stable complexes. These complexes have always a specific biological function; thus, it is important to identify and characterize the interaction between molecules, either DNA/RNA, DNA/DNA, protein/DNA, protein/protein, and so on. polycomb group proteins (PcG proteins) are epigenetic repressors involved in important physiologic processes as development and differentiation. They act on the chromatin through the formation of a repressive environment involving histone modification, recruitment of co-repressors, and chromatin–chromatin interactions. PcG form multiprotein complexes, whose characterization required several approaches. In this chapter, I will describe the co-immunoprecipitation (Co-IP) protocol, an easy method used to identify and analyze multiprotein complexes. In Co-IP, an antibody is used to isolate its target antigen, along with its binding partners, from a mixed sample. The binding partners purified with the immunoprecipitated protein can be identified by Western blot or by mass spectrometry.
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Lo Sardo, F. (2023). Co-Immunoprecipitation (Co-Ip) in Mammalian Cells. In: Lanzuolo, C., Marasca, F. (eds) Polycomb Group Proteins. Methods in Molecular Biology, vol 2655. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3143-0_6
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DOI: https://doi.org/10.1007/978-1-0716-3143-0_6
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