Recombinant full-length human NUAK2 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. NUAK2 or SNF1/AMP kinase-related kinase (SNARK) is a member of the NUAK family of SNF1-like kinase 2. NUAK2 is activated by muscle contraction and is a unique mediator of contraction-stimulated glucose transport in skeletal muscle (1). NUAK2 is involved in cellular stress responses linked to obesity and type 2 diabetes. NUAK2 shows kinase activity against a synthetic test peptide and the activity in keratinocytes is increased by AMP and 5-amino-4-imidazolecarboxamide riboside, implying that AMPK kinase-dependent pathway can activate NAUK2. Glucose deprivation also increases NAUK2 activity in baby hamster kidney fibroblasts.
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 chemical compounds have 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.
Kinase Enzyme System contains:
Kinase: NUAK2, 10μg (Human, recombinant full length). MW: ~110kDa. Substrate: CHKtide (KKKVSRSGLYRSPSMPENLNRPR); derived from the human CDC25C protein isoform A (amino acid 205–225). Other: Reaction Buffer, DTT.
Profile More Compounds In-House: ADP-Glo™ Kinase Assay + Kinase Enzyme System is optimized so that you are up and running in no time. Complete Systems: The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer, DTT and supplemental reagents as needed. Obtain Reliable Results: The broad dynamic range, the ease of use and better sensitivity obtained with ADP-Glo™ Kinase Assay result in less ambiguous data.
Notes
Kinase Enzyme System manufactured by SignalChem.
Bulk quantities available upon request.
References
1.Lefebvre, D.L. et al. (2001) Identification and characterization of a novel sucrose-non-fermenting protein kinase/AMP-activated protein kinase-related protein kinase, SNARK. Biochem. J. 355, 297–305.
2.Rune, A. et al. (2009) Regulation of skeletal muscle sucrose, non-fermenting 1/AMP-activated protein kinase-related kinase (SNARK) by metabolic stress and diabetes. Diabetologia. 52, 2182–9.
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