Recombinant human HIPK1 (156–555) was expressed in Sf9 cells using an N-terminal GST tag. HIPK1 or homeodomain-interacting protein kinase 1 is a ser/thr protein kinase and a member of the HIPK family. HIPK 1 is a nuclear kinase that phosphorylates homeodomain transcription factors. HIPK1 phosphorylates DAXX, and this leads to its relocalization and subsequent decrease in transcriptional repression activity (1). HIPK1 also interacts with p53 and phosphorylates it on serine residues. HIPK1 expression is elevated in breast cancer cell lines and embryonic fibroblasts from HIPK1-null mice showing more susceptibility to apoptosis induced by DNA damage (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.
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.Ecsedy, J.A. et al. (2003) Homeodomain-interacting protein kinase 1 modulates Daxx localization, phosphorylation, and transcriptional activity. Mol Cell Biol. 23, 950–60.
2.Kondo, S. et al. (2003) Characterization of cells and gene-targeted mice deficient for the p53-binding kinase homeodomain-interacting protein kinase 1 (HIPK1). Proc Natl Acad Sci USA 100, 5431–6.