Recombinant human RIPK1 (1-327) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. RIPK1 or Receptor Interacting Protein Kinase 1 is a serine/threonine kinase that was originally identified as interacting with the cytoplasmic domain of FAS. RIPK1 has been deemed as an important element in the signal transduction machinery that mediates programmed cell death. RIPK1 has been shown to interact with TRADD, TRAF1 TRAF2 and TRAF3 and TRADD can act as an adaptor protein to recruit RIPK1 to the TNFR1 complex in a TNF-dependent process. TNFα is capable of activating the noncanonical NF-κB pathway, but this activation of this pathway is negatively regulated by RIPK1. ADP-Glo Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects chemical compounds have on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.
