Webster Joshua D, Solon Margaret, Haller Susan, Newton Kim
Department of Pathology, Genentech, South San Francisco, CA, USA.
Department of Physiological Chemistry, Genentech, South San Francisco, CA, USA.
Methods Mol Biol. 2018;1857:153-160. doi: 10.1007/978-1-4939-8754-2_15.
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.
激酶RIPK3（受体相互作用蛋白激酶3）的激活是细胞发生坏死性凋亡的标志。RIPK3使自身和假激酶MLKL（混合谱系激酶样蛋白）磷酸化，导致MLKL易位至细胞膜并引起细胞裂解。因此，识别RIPK3自身磷酸化或MLKL上RIPK3依赖性磷酸化位点的抗体已被用于监测坏死性凋亡的诱导。在此，我们描述了对自磷酸化小鼠RIPK3进行免疫组织化学标记，作为检测小鼠组织中发生坏死性凋亡细胞的一种方法。
