NanoBRET™ TE Intracellular Kinase Cell-Based Assay Services
Quantifying kinase inhibitor occupancy, selectivity, affinity within the cellular environment where engagement would naturally occur is crucial to more accurately predict its engagement potencies against human kinases. In addition to equilibrium evaluation, it's also desirable to obtain kinetic parameters such as Residence Time for better compound optimization. Quantitative and wide-spectrum kinase profiling services using the NanoBRET™ Target Engagement Intracellular Kinase Assay System (Promega) enable you to assess your compound's engagement for a selected intracellular target under physiological conditions, including compound Residence Time at the target while keeping the cells intact.
Submit your compound(s) of interest, and Carna will rapidly deliver cellular IC50・% inhibition values and Residence Time!
Submit your compound(s) of interest, and Carna will rapidly deliver cellular IC50・% inhibition values and Residence Time!
Investigate compound functions in intact cells expressing full-length kinases
One type of tracer covers a wide-range of kinase investigations
Services
% inhibition measurements
(2 serial full-log dilutions,
3 conc. points in singlicate/duplicate)
(2 serial full-log dilutions,
3 conc. points in singlicate/duplicate)
IC50 determinations
(7 serial half-log dilutions, 8 conc.points in singlicate/duplicate)
(7 serial half-log dilutions, 8 conc.points in singlicate/duplicate)
Residence Time Analysis
(performed in triplicate based upon
your IC50 evaluation result)
(performed in triplicate based upon
your IC50 evaluation result)
IC50 Data Sample
Residence Time Analysis Sample
Improved selectivity under cellular ATP environment
[Target occupancy with 1μM Crizotinib]
(Right) using intracellular kinases in HEK293
(Left) using recombinant kinases assayed biochemically at Km ATP on Carna's MSA
NanoBRET™ System
This proximity-based assay detects protein interactions in live cells by measuring energy transfer from a bioluminescent protein donor (NanoLuc® luciferase) to a fluorescent probe (NanoBRET™ tracer). The NanoBRET™ TE Assay measures the apparent affinity of test compounds by competitive displacement of the NanoBRET™ tracer, reversibly bound to a NanoLuc® luciferase-kinase fusion protein expressed in live cells.
Please contact us for more information or if your target of interest is not included in the list.
A cell-permeable fluorescent NanoBRET™ tracer, a BRET acceptor, is added to HEK293 cells expressing a full length kinase/NanoLuc® fusion protein. Engagement of the tracer to the target protein generates a BRET signal
Binding of the test compound to the target protein results in a loss of NanoBRET™ signal between the target protein and the tracer inside intact cells
