MTase-Glo™ Methyltransferase Assay

Description

The MTase-Glo™ Assay is a bioluminescence-based assay that can be used to monitor the activities of methyltransferases (MTases) and their modulation by small molecules in a wide range of plate formats. The assay is well suited for high-throughput screening applications. The assay monitors formation of the reaction product S-adenosyl homocysteine (SAH) and can detect changes in activity of a broad range of methyltransferases, including DNA, protein, RNA and small molecule methyltransferases. The MTase-Glo™ Assay can be used for all classes of protein methyltransferases (lysine and arginine) and with different types of substrates (peptides, large proteins and even nucleosomes) to determine the specificity of these enzymes and their substrate requirements.

After the methyltransferase reaction is complete, the MTase-Glo™ Reagent is added to convert SAH to ADP. The MTase-Glo™ Detection Solution is then added to convert ADP to ATP, which is detected via a coupled luciferase reaction (Figure 1). Luminescence is measured using a plate-reading luminometer and can be correlated to SAH concentration using an SAH standard curve (Figure 2). The half-life of the luminescent signal is greater than 4 hours. This extended signal half-life eliminates the need for luminometers with injectors and allows batch-mode processing of multiple plates.

Features – Benefits

• Easily Monitor Methyltransferase Activity : Simple add-and-read format makes it easy to monitor methyltransferase activity.
• Use Any Methyltransferase : Can be used with any methyltransferase that uses S-adenosyl methionine (SAM) as the methyl group donor.
• Experience Low False Hits : Bioluminescent assay optimized for screening applications; no concerns about fluorescence interference.
• Use Less Enzyme : Low background and large dynamic range of assay produces excellent signal-to-noise ratios at low enzyme concentrations.
• Use Natural Substrates : No need to modify substrates, which can lead to kinetic artifacts.
• Enjoy Flexibility : No interference from high concentrations of SAM in assay.

Applications

• High-throughput systems.