The NanoBRET assay provides real-time data of the binding affinity of a compound to kinase target including occupancy time Intact cells provide a relevant environment for the compound-kinase interaction in regards of ion and ATP concentration, pH, or co-factors. In addition, to function inside the cell, compounds must overcome the cellular membrane. Suitable for high-throughput screening Kinase activity testing in intact cells can be investigated with our Cellular Phosphorylation Assay
Setups: IC50 value determination in 10-dose duplicate assay format. Free choice for screening of the kinase targets of your choice. New kinase target development/validation is available upon request.
Controls: We use DMSO as a negative control. In every project, testing of a standard reference compound is included in IC50 assay format as well.
Turnaround time: 2-3 weeks. Expedited scheduling and data delivery can be arranged prior to study commencement. Report: A detailed report including assay conditions and comprehensive evaluation of data as well as raw data for each analysis will be provided.
Compound requirements: In brief, 50 to 100 µl of a 10 to 50 mM DMSO stock or 2-3 mg solid material is needed.
Please refer to our FAQs for information regarding compound preparation and shipping.
Please download our assay condition form.
Purpose: Testing the efficacy of epigenetic drug candidates to interrupt the interaction of a bromodomain protein with chromatin
Targets: Bromodomains
Assay Format: NanoBRET Bromodomain: Histone Interaction Assay (Promega)
HEK 293T cells are transfected with two constructs: a bromodomain-NanoLuc fusion construct and a Histone-Halo Tag fusion construct. After 24 hours, the Halo Ligand and the test compound will be added. The interaction of bromodomain and histone will allow for electron transfer between the NanoLuc to the Halo Ligand eliciting fluorescence in the presence of the NanoLuc substrate.
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