FLT3 (D835Y) 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 IKKβ Kinase Enzyme System contains:

• IKKβ Kinase, 10μg (Human, recombinant full-length). MW: ~105kDa.
• IKKtide (KKKKERLLDDRHDSG-LDSMKDEE) Substrate; derived from human IkBA (amino acids 2141).
• Reaction Buffer, DTT.

Full-length recombinant human IKKβ was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. IKKβ is a serine/threonine protein kinase that phosphorylates the I-kappa-B protein, which is an inhibitor of the transcription factor NF-kappa-B complex. Phosphorylation of I-kappa-B triggers the degradation of the inhibitor via the ubiquitination pathway, thereby activating NF-kappa-B complex. The activity of IKKβ is stimulated by TNF and IL1 and IKKβ forms a heterodimer that interacts with NIK. Overexpression of catalytically inactive IKKβ blocks cytokine-induced NF-kappa-B activation. Aspirin and sodium salicylate can specifically inhibit IKKβ activity in vitro and in vivo by binding to IKKβ to reduce ATP binding.

IKKβ NCBI Database Entry.

The IKKβ 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.

See all Kinase Enzyme Systems available from Promega.