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Kinases

Kinase enzymes are at the heart of cellular communication, playing a pivotal role in translating external signals into functional responses within cells. Their involvement in cell growth, division, and death makes them key targets in the development of therapies for diseases such as cancer, where these processes are dysregulated. The need to understand 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 addition of reagents, substrates, and inhibitors at specific concentrations, volumes, and times. The Myra liquid handling system has been 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 kinase large molecule high throughput screening for drug discovery.

Image courtesy Invitrogen Corporation, Walthem MA.

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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ADPGlo Assay

Case Study: Promega ADP-Glo Kinase Assay

Reliable, Reproducible Kinase Inhibitor Screening

ADP-Glo Kinase time course assay was run to determine the enzyme kinetics of a kinase with a peptide substrate.

In a typical ADP-Glo™ Kinase Assay, the process starts with the kinase reaction, where a kinase enzyme phosphorylates a substrate in the presence of ATP. Following this, the ADPGIo™ Reagent is added to stop the kinase reaction and deplete remaining ATP, preparing the system for ADP measurement.

Finally, the Kinase Detection Reagent converts ADP back to ATP, which is quantified through a luminescent signal.

Automating this sequence with Myra ensured that each step was precisely timed and executed, maintaining the integrity and consistency of the assay across multiple wells and plates.

Download Case Study PDFView Scripts

Experimental Setup

Small volumes done really well. 

DP-Glo Kinase Assay was conducted using peptide diluted 1:2 in a total volume of 2 mL. The peptide was diluted across the plate followed by the addition of 2 mL of kinase. Four microliters of a stop solution were added at 5 min intervals from time zero.

The systematic addition of components needed to take into account the specific times required to ensure accurate addition of the stop solution. An ADP standard curve was created in triplicate to allow for ATP to ADP conversion calculations.

Results

High quality data.

Myra facilitates the generation of data sets necessary for Michaelis-Menten analysis by ensuring that reaction conditions are consistently replicated. This uniformity allows for the precise determination of reaction velocities at varying substrate concentrations, critical for plotting Michaelis-Menten curves and calculating kinetic parameters.

Kinase large molecule high throughput screening on Myra enables rapid and reproducible analysis of numerous inhibitors, accelerating the identification of promising therapeutic candidates. Integrating with a q-PCR Machine further supports downstream validation, connecting biochemical screening data to gene expression or cellular outcomes.

Figure 1. CHK1 kinase activity was assessed using the CHK1 Kinase Enzyme System in combination with ADP-Glo, following treatment with a 22-point, 2-fold serial dilution of Staurosporine (blue circles) and AZD7762 (red squares). Data represent the mean ± standard deviation of technical replicates.

Frequently Asked Questions

What are kinase assays used for?
Kinase assays help researchers understand kinase‑mediated signalling and support the discovery of therapeutics. Assays are especially useful in oncology, where abnormal kinase activity drives disease progression.
How does Myra improve the accuracy and reliability of kinase assays?
Myra combines closed‑loop 100 μm position control with an integrated camera to deliver 5 % CV precision at 1 µL and 1 % CV from 5–50 µL, ensuring reproducible low‑volume dispensing and minimising human error.
What advantages does TR‑FRET offer in kinase assay applications?
The TR‑FRET format minimizes background fluorescence and maximizes signal‑to‑noise, providing a sensitive, high‑throughput readout for protein interaction studies in drug discovery screens.
Why are kinase assays important in drug discovery?
Kinases regulate cell growth, division, and death. Accurately measuring their activity guides the design and optimisation of candidate inhibitors during early‑stage drug discovery.
What are the benefits of automating kinase assays using Myra?
Automating reagent additions with Myra reduces manual intervention, cuts the risk of pipetting errors, and enables fast, consistent screening of kinase inhibitors across multi‑well plates.
Is Myra suitable for research labs as well as pharmaceutical screening?
Yes. Myra is trusted by labs across the US, Europe, Africa, South America, and Asia, delivering the same precision platform for small exploratory studies and large‑scale screens.
Where can I learn more?
Request a demo or reach out to our applications team with any questions.