Detection of necroptosis by phospho-MLKL immunohistochemical labeling

TNF-induced necroptosis is involved in many physiological and pathological processes. Phospho-MLKL is a hallmark of necroptosis. Cecum is a sensitive organ with extensive necroptosis responses to TNF in vivo. Here, taking advantage of commercially available mouse TNF and easily accessible reagents and materials, we systematically provide a detailed and highly versatile protocol of detecting necroptosis signaling in mouse cecum by immunohistochemical labeling, which can also be used in other tissues or antibodies in immunohistochemical staining. For complete details on the use and execution of this protocol, please refer to Yang et al. (2020) and Chen et al. (2015).