Kinase RR kit - The Kinase RR (Reaction Rate) kit enables easy and reliable identification of protein kinase inhibitors form large compound libraries. The kit uses direct bioluminescent measurement of the ATP degradation rate to distinguish between hits and non-hits. The kit allows 1000 determinations using 96 well plates or 2000 using 384 well plates.
ATP Kit SL - Any enzyme or metabolite participating in an ATP forming or degrading reaction can be monitored using ATP technology. Adding ATP Reagent SL to a sample makes it possible to continuously monitor the ATP level by measuring the intensity of the emitted light .
Kinase RR Kit or Kinase Reaction rate Kit is developed for the easy and reliable identification of protein kinase inhibitor candidates from large compund libraries. The Kinase RR Kit uses direct bioluminescent measurement of ATP degradation rate to distingusih hits from non-hits. In contrast to other bioluminescent ATP based assays of kinase there is no interference from inhibitors acting on luciferase. Furthermore the standard curve is linerar rather than sigmodial. the Kinase RR Kit is flexible and can easily be adapted to various HTS strategies. Z’ values up to 0.96 have been reported. The assay does not require radioactive materials, antibodies or any form of conjugates.
Applications - Kinase RR Kit is based on a method of measuring the first order reaction rate constant of protein kinase reactions by monitoring the ATP degradation rate using bioluminescence. The assay starts by adding ATP to a reaction mixture containing all necessary assay components, including luciferase and luciferin. In HTS the light is measured at two points in time. In non-HTS the light can be measured repeatedly until a significan reduction of light is obtained. The assay is set up so that a) the kinase reaction follows first order kinetics, b) the sample light emission, normalized against the blank (no kinase), is proportional to the remaining ATP concentration.
The first order rate constant of the kinase reaction is proportional to the kinase activity over several orders of magnitude, which makes it very easy to set up new assays. Furthermore variation in ATP and luciferase cancel out in calculations and do not affect assay results.
Instruments - It is recommended to use a microplate luminometer with a dispenser for adding ATP. However, microplate luminometers without dispensers can be used. In such case please consult BioThema AB for technical support.
Assay procedure - Detailed procedures on Standard Curve Generation, Substrate Optimisation, HTS and IC50 Determination are found in the “Instructions for Use”, which is included with every kit.
ATP Kit SL - With ATP reagent SL you get a stable light, which makes it possible to perform kinetic as well as end-point assays. At the end of the assay a known amount of ATP is added (internal ATP Standard) to the reaction mixture for calibration and to calculate the formation or degradation of ATP in moles.
Applications include lipolysis, oxidative phosphorylation and continous monitoring of cell lysis.
If no immediately obvious reaction can be coupled to ATP formation or degradation, e.g. pyrosequencing or measurement of DNA polymerase, please contact us at email@example.com for recommendations of assay development.