Description
A Kinase-NanoLuc® fusion vector is introduced by transfection into mammalian cells and allowed to express. As NanoLuc® luciferase is extremely bright, only low expression of the kinase-NanoLuc® fusion protein is needed.
In the NanoBRET® TE Kinase Assay, a cell-permeable fluorescent NanoBRET™ tracer, a NanoLuc® substrate, and the Extracellular NanoLuc® Inhibitor are supplied. Addition of these to cells expressing the kinase-NanoLuc® fusion allows a strong BRET signal to be achieved between the kinase-NanoLuc® protein and NanoBRET™ tracer. The extracellular NanoLuc® inhibitor ensures that the BRET signal achieved is from live, uncompromised cells.
The presence of unlabled test compound that binds to target kinase results in a loss of BRET signal. As BRET has tight distance constraints, the data obtained is specific for the kinase fused to NanoLuc® luciferase. In addition, the data results in a quantitative intracellular affinity provided the appropriate tracer concentration is used.
PRODUCTS
- Intracellular TE Nano-Glo® Substrate/Inhibitor
- Intracellular TE Nano-Glo® Vivazine™/Inhibitor
- CC1 pan-Kinase Inhibitor
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