RIPK1 Kinase Enzyme System

The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The RIPK1 Kinase Enzyme System contains:

• RIPK1 Kinase, 10μg (Human, recombinant; amino acids 1–327); MW: ~63kDa
• Native Swine Myelin Basic Protein (MBP) Substrate, (1mg/ml)
• 5X Reaction Buffer;  MnCl₂, 2.5M; DTT, 0.1M

Recombinant human RIPK1 (amino acids 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.

Application Note 

NCBI Database Entry

The RIPK1 Kinase Enzyme System can be purchased with or without the ADP-Glo™ Kinase Assay reagents. Used together, the ADP-Glo™ Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.

Use with ADP-Glo™ Kinase Assay

The 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 of chemical compounds 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.