Purification and analysis of cell death complexes

Cell death complexes, such as the complex II and necrosome, play pivotal roles in executing programmed cell death pathways. Understanding the composition and dynamics of these complexes is essential for elucidating the molecular mechanisms underlying apoptosis and necroptosis. This protocol describes a robust method for the affinity purification of endogenous or tagged cell death complexes from mammalian cells, followed by analysis through western blotting and optional mass spectrometry. This approach enables investigation of protein-protein interactions and the dynamic assembly of complexes under various stimuli or genetic conditions, providing insights into their mechanisms in health and disease.