IKK beta Kinase Enzyme System

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 IKK? Kinase Enzyme System contains:

• IKK? Kinase, 10ug (Human, recombinant full-length). MW: ~105kDa.
• IKKtide (KKKKERLLDDRHDSG-LDSMKDEE) Substrate; derived from human IkBA (amino acids 21–41).
• Reaction Buffer, DTT.

Full-length recombinant human IKK? was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. IKK? 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 IKK? is stimulated by TNF and IL1 and IKK? forms a heterodimer that interacts with NIK. Overexpression of catalytically inactive IKK? blocks cytokine-induced NF-kappa-B activation. Aspirin and sodium salicylate can specifically inhibit IKK? activity in vitro and in vivo by binding to IKK? to reduce ATP binding.

IKK? NCBI Database Entry.

The IKK? 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.