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Cambrex Corporation
The PKLight™ HTS Protein Kinase Assay Kit is a patented generic, homogeneous assay designed for use in high throughput screening of kinase activity. The kit contains all reagents necessary for the assay and has been designed for use in a cell free system with either 96, 384 or 1536 well microtiter plates. All that is required from the user is a purified form of the kinase of interest, a suitable substrate, buffer and ATP. The PKLight Assay can be easily optimized for each kinase/substrate pair to produce rapid, reproducible data suitable for IC50 determinations of screen compounds. This technology does not require radioactive beads, radio-labeled ATP, specially designed antibodies or specifically modified substrate sequences.
The PKLight Assay exploits the kinases’ intrinsic ATPase activity, resulting in the cleavage of the gamma-phosphate moiety of ATP and its subsequent insertion into the target substrate. This results in the phosphorylation of the substrate and the conversion of ATP to ADP. The PKLight Assay measures the consumption of ATP and is based upon the bioluminescent measurement of the remaining ATP present in the wells after the kinase reaction.
PKLight is a Protein Kinase Assay. The assay and kits are protected by UK patent number GB 2,375,171 B, US patent number 6,599,711 and International Patent Application number PCT/GB01/05506.
* Generic assay platform – single endpoint for all kinase/substrate pairs.
* Simple, homogeneous assay – following the kinase reaction, simply add a single reagent and read in a plate luminometer.
* Wide dynamic range – assay is linear over a wide ATP concentration range, 0-20 µM.
* Highly sensitive – assay can be used with small amounts of kinase enzymes and substrates.
* Fast results – measurement times can be less than 1 second per assay point.
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