NEK3 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 NEK3 Kinase Enzyme System contains:

• NEK3 Kinase, 10μg (Human,recombinant full length). MW: ~86kDa.
• Native Swine Myelin Basic Protein (MBP) Substrate.
• Reaction Buffer, DTT.

Recombinant full-length human NEK3 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. NEK3 is a member of the NEK family of protein kinases that share high amino acid homology with NIMA (never in mitosis gene a). NEK3 mRNA is detected in all the proliferating cell lines with the amount not changing during the cell cycle. During Prolactin receptor signaling, VAV2 is phosphorylated and activated by NEK3. Overexpression of NEK3 in Chinese hamster ovary cells increases cytoskeletal re-organization in response to Prolactin while downregulation of NEK3 expression by siRNA blocks these effects. Prolactin also stimulates interaction between NEK3 and paxillin leading to increased paxillin phosphorylation. Analysis of breast tissue microarrays show a significant up-regulation of NEK3 expression in malignant versus normal specimens.

NEK3 NCBI Database Entry.

The NEK3 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.