Recombinant human MRCKalpha (1–473) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Myotonic Dystrophy Kinase-Related Cdc42-Binding Kinase α (MRCKα) is a Cdc42/Rac/Rho interactive/binding serine/threonine kinase with multiple functional domains (1). MRCKs are effectors of RhoA and Cdc42, respectively, for actin reorganization. MRCKα is a critical regulator of signal transduction pathways in eukaryotic cells that are known principally for their role in regulating the cytoskeleton, and they do so by recruiting a variety of downstream effector proteins (2).
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: MRCKα, 10μg (Human, recombinant; amino acids 1–473). MW: ~73kDa. Substrate: S6K substrate (KRRRLASLR); derived from human 40S ribosomal protein S6 (amino acid 230–238). 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.Ivan, T. et al. (2003) Genomic organization of human myotonic dystrophy kinase-related Cdc42-binding kinase α reveals multiple alternative splicing and functional diversity. Gene 304, 107–115.
2.Ivan, T. et al. (2001) Phosphorylation of a novel myosin binding subunit of protein phosphatase 1 reveals a conserved mechanism in the regulation of actin cytoskeleton. J. Biol. Chem. 276, 21209–16.
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