Detection of necroptosis by phospho-RIPK3 immunohistochemical labeling

Abstract

Activation of the kinase RIPK3 (receptor interacting protein kinase 3) is a hallmark of cells dying by necroptosis. RIPK3 phosphorylates both itself and the pseudokinase MLKL (mixed lineage kinase-like) resulting in MLKL translocation to membranes and cell lysis. Antibodies recognizing RIPK3 autophosphorylation or the RIPK3-dependent phosphorylation sites on MLKL have therefore been used to monitor necroptosis induction. Here we describe immunohistochemical labeling for autophosphorylated mouse RIPK3 as a means of detecting cells undergoing necroptosis in mouse tissues.