title: Activity assays for receptor-interacting protein kinase 1:a key regulator of necroptosis.
abstract: Necroptosis is a novel form of regulated non-apoptotic cell death, which displays morphological features of necrosis. The kinase activity of receptor-interacting protein kinase 1 (RIP1) is a critical component in signaling for necroptosis. The development of assays to evaluate RIP1 kinase activity is important in the further development of existing and novel inhibitors of necroptosis. Here, we describe RIP1 protein expression and purification from mammalian and insect cells as well as two in vitro kinase assays to detect RIP1 kinase activity and inhibition.
authors.affiliation: Department of Biochemistry, School of Medicine, Tufts University, Boston, MA, USA.
authors.forename: Jenny L
authors.initials: JL
authors.lastname: Maki
authors.forename: Alexei
authors.initials: A
authors.lastname: Degterev
mesh_terms: D000818:Animals; D057075:Enzyme Assays / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism; D007211:Indoles / Q000737:chemistry / Q000378:metabolism; D009336:Necrosis / Q000473:pathology*; D011874:Radiometry; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification; D061987:Sf9 Cells
chemical_list: D007093:Imidazoles; D007211:Indoles; D011994:Recombinant Proteins; C507699:necrostatin-1; D053422:Receptor-Interacting Protein Serine-Threonine Kinases