Recombinant full-length human CDC7 and DBF4 proteins were co-expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. CDC7 is a cell division cycle 7 homolog protein that is critical for the G1/S transition and is also essential for initiation of DNA replication as cell division occurs. CDC7 is expressed in many normal tissues, but the overexpression of CDC7 may be associated with neoplastic transformation for some tumors and transformed cell lines (1). CDC7/DBF4 kinase promotes S phase by alleviating an inhibitory activity in Mcm4 that evolved to integrate several protein kinase (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: CDC7/DBF4 , 10μg (Human, recombinant full-length). MW: ~94kDa (CDC7) and ~125kDa (DBF4). Substrate: PDKtide (KTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC); derived from two human proteins: residues 1–14 are based on AKT1 (307–320), and residues 16–39 are based on PKN2/PRK2 (961–984). 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.Hess, G.F. et al. (1998) A human homolog of the yeast CDC7 gene is overexpressed in some tumors and transformed cell lines. Gene 211, 133–40.
2.Sheu, Y.-J. et al. (2010) The Dbf4-Cdc7 kinase promotes S phase by alleviating an inhibitory activity in Mcm4. Nature 463, 113–7.
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