tyrosine protein kinase assay

Protein kinases are recognised as important drug targets due to the crucial role they play in human diseases such as cancer, inflammatory diseases, neuronal and metabolic disorders. Identification of protein kinase modulators has become a common strategy for drug discovery.

The FLEXYTE® lifetime tyrosine protein kinase assay utilises a unique long lifetime fluorophore coupled to a panel of generic and bespoke peptide substrates to provide a homogeneous, robust, antibody-free and cost-effective screening solution. When analysed with TTP Labtech’s ameon® lifetime reader the resultant platform offers intrinsic tolerance to common assay interferences to deliver unparalleled assay robustness.

assay principle

Tyr_Protein_Fig1

 

Fig 1. FLEXYTE lifetime tyrosine protein kinase assay

benefits of FLEXYTE lifetime assays

The FLEXYTE lifetime tyrosine protein kinase assay platform has been developed through the design and synthesis of peptide substrates labelled with the long lifetime FLEX17 fluorophore. The presence of the native tyrosine residue in the substrate sequence reduces the lifetime of the fluorophore. Phosphorylation of this tyrosine by a corresponding kinase alleviates this effect leading to an increase in lifetime which correlates with the level of phosphorylation of the substrate (Fig 1).

Utilisation of long lifetime FLEX17 fluorophore enables the discrimination of substrate and product from fluorescent interfering compounds whose lifetimes are commonly below 2 nanoseconds. It also provides a stable, defined assay window upon which assays can be built in 384 or 1536-well microplates. FLEXYTE tyrosine kinase assays can be run in real-time or in a stopped format depending on end-user requirements.

ameon lifetime reader

TTP Labtech’s ameon lifetime reader meets the challenge of accelerating the drug discovery process by integrating easily into screening workflows and offering a reliable fluorescence lifetime readout. The instrument applies a proprietary data recording technology to perform realtime decay curve capture and delivers precise lifetime calculation at high throughput.

specific enzyme list

Bespoke FLEX17-labelled peptides can be configured to meet the requirements of many tyrosine protein kinases.

Tyr_Protein_Fig2

View our tyrosine protein kinase assays app note

 

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