Application note: Promega’s ADP-Glo™ assay
This application note discusses how the ADP-Glo™ Kinase assay is a universal, homogeneous, high-throughput screening method that measures kinase activity by quantifying the amount of ADP produced during the kinase reaction.
Kinases are a large and diverse group of enzymes that are involved in many cellular metabolic and regulatory processes. During the kinase reaction, substrates are phosphorylated while ATP is converted into ADP. Due to the kinases’ pivotal role in intracellular signaling they are prominent drug targets.
The ADP-Glo™ Kinase assay is a universal, homogeneous, high-throughput screening method to measure kinase activity by quantifying the amount of ADP produced during the kinase reaction. It first eliminates remaining ATP and secondly converts ADP into ATP. The ATP is then quantified in an ATP-dependent light-emitting luciferase reaction.
To mimic an enzymatic reaction, an ADP/ATP standard curve was prepared from the nucleotide stock solutions that were supplied with the kit. Measuring the assay on a BMG LABTECH microplate reader revealed a signal stabilization around 20 min and that the instrument gives a linear signal from 1 mM down to >0.1 µM ADP.
This application note is restricted - login or subscribe free to access
Thank you for visiting our website. To access this content in full you'll need to login. It's completely free to subscribe, and in less than a minute you can continue reading. If you've already subscribed, great - just login.
Why subscribe? Join our growing community of thousands of industry professionals and gain access to:
- quarterly issues in print and/or digital format
- case studies, whitepapers, webinars and industry-leading content
- breaking news and features
- our extensive online archive of thousands of articles and years of past issues
- ...And it's all free!
Related content from this organisation
- Application note: Screening for optimal association and dissociation rates of ligands using a live cell binding assay
- Expert view: Promising methods in kinase screening
- Screening In-Depth Focus 2019
- Under the microscope: Improving hit discovery efficiency at Europe´s leading screening centre
- Webinar highlights: Cell-based and biochemical high-throughput screening in 1536-well plates