Kinases
Kinase enzymes are at the heart of cellular communication, playing pivotal roles in translating external signals into functional responses within cells. Their crucial involvement in cell growth, division, and death makes them key targets in the development of therapies for diseases where these processes are dysregulated, such as cancer. The need to understand and manipulate kinase activity for therapeutic purposes has given rise to the development and refinement of kinase assays in drug discovery.
Accurate measurement of kinase activity depends on the meticulous addition of reagents, substrates, and inhibitors at specific concentrations, volumes, and times. Myra has shown to provide unmatched precision in liquid dispensing, reduced manual intervention, and minimized human errors, revolutionizing the conduct of kinase assays.
ADP-Glo
The ADP-Glo Kinase Assay is a luminescent assay that measures the amount of adenosine diphosphate (ADP) produced by kinase reactions over time.
It is used to evaluate the activity of kinase enzymes and to screen for kinase inhibitors by quantifying the conversion of ATP to ADP in a time-course manner, making it a valuable tool for drug discovery and development programs targeting kinase-driven pathways.
TR-FRET
Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) is a sensitive and reliable fluorescence-based assay technique that combines the principles of time-resolved fluorescence and FRET to minimize background signal and maximize signal-to-noise ratio. It is widely used in biochemical and biological research for studying molecular interactions, such as protein-protein or protein-DNA interactions, enabling the detection and quantification of these interactions in various assay formats, including high-throughput screening for drug discovery.
Kinase Assays on Myra
Browse a comprehensive table of all available templates on Myra, complete with direct links to specific MyraScript pages. Access protocol documentation via a dedicated PDF link. Verified options include templates for BMS, Partner (kit company), and the Myra community (customers).
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Case Study: Promega ADP-Glo Kinase Assay
Reliable, Reproducible Kinase Inhibitor Screening
High-throughput screening of kinase inhibitors demands both precision and consistency, particularly when working at low volumes. In this case study, Myra was used to automate the ADP-Glo™ Kinase assay, demonstrating its ability to perform complex pipetting routines with exceptional accuracy.
Even across a 22-point dose–response series, results showed minimal variability and strong agreement with published IC₅₀ values, confirming Myra’s suitability for compound screening and kinetic analysis. This study highlights Myra’s strength in delivering reproducible, high-quality data in demanding drug discovery workflows.
Experimental Setup
Small volumes done really well.
CHK1 kinase activity was assessed using an ADP-Glo kinase assay, following a 1 mL treatment with a 22-point, 2-fold serial dilution of Staurosporine or AZD7762, run in duplicate. Reactions contained 2 mL of 4 ng CHK1, 2 mL of 50 μM ATP/0.2 μg/μL CHKtide Substrate and were incubated for 60 minutes at room temperature. Reactions were carried out on a 384-well flat-bottomed microplate.
Precise timers were integrated into the script for the systematic addition of Stop and Detection reagents ensuring kinetic consistency across the plate. Fluorescence was read on a Hidex plate reader.
Results
High quality data.
Relative luminescence units (RLU) were plotted against log10-transformed inhibitor concentrations (nM). A four-parameter logistic (4-PL) non-linear regression provided IC50estimates for Staurosporine and AZD7762.
Staurosporine and AZD7762 displayed distinct potencies, with IC50 values of 1.52 nM and 5.74 nM respectively, indicating that AZD7762 is approximately 3.8 times less potent under identical assay conditions. The resulting dose–response curves showed excellent fit (R2 > 0.99), and all values fell within 0.5 nM of published IC50 data, validating both assay fidelity and Myra’s pipetting precision.
Technical replicate variability remained below 8% across the entire concentration range, demonstrating the reliability of automated pipetting even at such low volumes.
