Current methods to assess kinase activity have been performed using radioactive isotopes or highly specific antibodies. Despite its accuracy and quality of results, the downsides to these methods are its cost and relative complexity both in preparation and result interpretation.
The IMAP TR-FRET Detection System is designed for its proprietary IMAP platform and offers numerous advantages as it is antibody independent and stable for greater than 24 hours.
With no antibodies, searches for a compatible antibody are eliminated accelerating the assay development process. Signal stability that is greater than 24 hours allows customers to schedule their screens compared to alternative TR-FRET detection methods that require more rigorous scheduling of screens.
IMAP FP allows greater substrate choice enabling researchers to work with larger substrates such as proteins. It also provides greater sensitivity, which reduces the amount of enzyme required and ultimately lowers the overall cost to the assay.
"The IMAP platform is becoming the preferred assay to screen kinase activity as it is applicable to all regions of the human kinome," said Susan Clark, director of Reagents Marketing at Molecular Devices.
"The IMAP platform enables our customers to develop assays for uncommon and proprietary enzymes with a homogeneous, antibody independent system," Clark continued.
IMAP is a technology based on the specific, covalent-coordinate, high-affinity interaction of trivalent metal containing nanoparticles with phosphogroups.
These phosphogroups can be free, linked to serines, threonines or tyrosines, or other molecules which make IMAP a generic platform to assess kinase, phosphatase and phosphodiesterase activity.
This basic principle has been used in the IMAP Binding System using both fluorescence polarisation and TR-FRET (as a read-out).
In a microwell assay format, fluorescently labelled peptides are phosphorylated in a kinase reaction.
Addition of the IMAP Binding System stops the kinase reaction and specifically binds the phosphorylated substrates. Phosphorylation and subsequent binding of the substrate to the beads can be detected either by FP or TR-FRET.
With the IMAP Platform binding systems, Substrate Finder Plates, validated substrates, and the FP and TR-FRET detection modes, transitions from assay development, screening and hit evaluation can be accelerated for virtually any kinase.
Kinases are implicated in many diseases, such as cancer, diabetes, cardiovascular disease, and neurological disorders.
It is estimated that kinase malfunction contributes to more than 400 diseases and that greater than 20 per cent of the drug discovery efforts are focused on protein kinase inhibitors.
Researchers require technologies that will enable them to rapidly develop assays that provide data for these potentially important kinase inhibitors.