Frequently Asked Questions
Some of the commonly asked questions about the technology, assay and sample submission are given below. If you have additional questions, please contact Luceome at 520-495-0161 or info@luceome.com.
KinaseSeeker Assays
KinaseSeeker is a binding assay based on a patented cell-free split-luciferase technology. Split luciferase fragments fused to a scaffold protein and kinase are translated in vitro in a cell-free system and reassembled into a functional luciferase in the presence of a kinase active site binding probe. KinaseSeeker assay has the following benefits over other assay techniques:
- Luminescence detection, hence low false positives from fluorescent compounds
- Binding assay, hence identification of active site directed and allosteric inhibitors is possible
- Cell-free format, hence ability to identify inhibitors in a complex cellular environment at cellular ATP concentrations
- Homogeneous assay, hence not affected by artifacts due to immobilization
- Sensitive with Low Background; Broad dynamic range
We currently have 415 protein kinases spanning all families on our panel. We are continuously working to add more kinases. If you would like to stay informed when new kinases are added, please send us an email at info@luceome.com to be added to our mailing list.
We offer an a la carte service, where you can build your own panel and profile kinases of your choice. In addition, we offer several preset panels, comprising popular kinases, which can be used to profile your compounds cost effectively.
Yes. We have high throughput capabilities for rapid evaluation of compound libraries against a target kinase.
Yes. KinaseSeeker is a luminescence based assay, which eliminates interference and false positives that result from fluorescent compounds.
We test all compounds for luciferase inhibition prior to testing for kinase inhibition. If your compound inhibits luciferase in our control assays, we will notify you ahead of initiating profiling experiments. These control experiments are part of the profiling services we provide for kinases. In the event that your compound cannot be tested for kinase inhibition, you will not be charged for the control experiments.
The KinaseSeeker assay entails incubation of the translated kinase with the inhibitor for 1 hour at room temperature in rabbit reticulocyte lysate. We have tested the concentration of ATP in several batches of lysate using Promega’s ENLITEN assay. The ATP concentration was approx. 1 mM, which is well above its Km for most protein kinases.
Upon completion of testing (~10-15 business days), an electronic copy of the study report is submitted to the customer via email. The screening data is reported as % activity remaining for each kinase and compound profiled, where a lower number indicates a more potent compound.
Yes. In addition to Type I inhibitors, KinaseSeeker can be used for detection of Type II and other allosteric inhibitors.
