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Encompassing Amino Acids: CDC7: 1-574(end), DBF4: 1-674(end)
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 5 mM MOPS, 2.5 mM β-glycerophosphate, 5 mM MgCl2, 1 mM EGTA, 0.4 mM EDTA, 0.05 mM DTT, 50 ng/ml BSA, 2 mM ATP, 200 µg of PDKtide synthetic peptide substrate. Add [33P]-ATP and incubate at 30°C for 15 minutes, then spot reaction on phosphocellulose paper, fix in 1% phosphoric acid, and assay with a scintillation counter.
Background: 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. CDC7/DBF4 kinase promotes S phase by alleviating an inhibitory activity in Mcm4 that evolved to integrate several protein kinase
Description: CDC7/DBF4 complex of human CDC7, also known as HsCDC7 and Hsk 1, GenBank Accession No. NM_003503 a.a. 1-574(end) with N-terminal GST-tag and MW= 94 kDa, and human DBF4, also known as ASK and CHIF, GenBank Accession No. NM_006716, a.a. 1-674(end) with N-terminal GST-tag and MW= 125 kDa, co-expressed expressed in an Sf9 infected baculovirus expression system.
Format: Aqueous Buffer Solution
Formulation: 50 mM Tris-HCL, pH 7.5, 150 mM NaCl, 10 mM glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF and 25% glycerol.
Genbank: CDC7: NM_003503.4; DBF4: NM_006716.4
Storage Stability: At least 6 months at -80°C.
Tags: CDC7: N-terminal GST-tag
DBF4: N-terminal GST-tag
Uniprot: O00311 (CDC7), Q9UBU7 (DBF4A)
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Hess, G. F. et. al. Gene 211: 133-140, 1998
2. Sheu, Y.-J et. al. Nature 463: 113-117, 2010
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