Description
Convenient, Scalable Kinase Profiling
The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The KSR1 (L639F) Kinase Enzyme System contains:
KSR1 (L639F) Kinase, 10μg (Human, recombinant; amino acids 335–end); MW: ~105kDa
CREBtide substrate (1mg/ml; KRREILSRRPSYR) derived from human CREB1 isoform A (amino acids 109–121)
5X Reaction Buffer; DTT, 0.1M
Recombinant human KSR1 (L639F) (amino acids 335–end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. KSR1 or Kinase Suppressor of Ras 1 interacts with various kinases of the Raf/MEK/extracellular signal-regulated kinase pathway to enhance its activation. KSR1 is regulated in response to a specific mode of dimerization of its kinase domain, which is termed the side-to-side dimer, whereas KSR1 also participates in forming side-to-side heterodimers with RAF and this can trigger RAF activation. KSR1 has an essential protective role in the intestinal epithelial cell during inflammation through activation of cell survival pathways.
The KSR1 (L639F) 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.
