EMD Millipore, the Life Science division of Merck KGaA of Darmstadt, Germany, announced the launch of the SIRTainty Class III HDAC Assay for detection of sirtuin activity with increased sensitivity, flexibility, and reliability.
The SIRTainty assay utilizes a novel, patent-pending technology for the sensitive detection of all known sirtuin family members. Unlike conventional assays that are dependent on a single, pre-labeled fluorescently tagged substrate, this new assay employs untagged acetylated peptide substrates. This design enables unparalleled flexibility in the choice of sirtuin isoform and peptide substrate, and eliminates the potential for artifacts resulting from use of artificial substrates containing bulky fluorophores.
The SIRTainty assay is ideal for a range of applications including:
• Assays of peptide substrate panels to determine enzyme-substrate specificity
• Screening for sirtuin activators and inhibitors and assessment of potency
• Activity comparisons of sirtuin family members
• Enzyme kinetics studies
The assay offers a limit of detection as low as 0.16U of purified sirtuin, a nearly five-fold greater sensitivity than assays dependent on a fluorescently labeled substrate. This homogenous, no-wash, 96-well format minimizes hands-on time, speeds the overall workflow and can be completed in 90 minutes.
The SIRTainty assay allows researchers to reliably measure sirtuin activity using a wide variety of substrates and also greatly facilitates the discovery and characterization of sirtuin modulating compounds. Previous assays have employed artificial substrates bearing large, bulky fluorophores, not found within the cell. This complication has made it difficult to translate in vitro findings on activators/inhibitors to a cellular context. Because the SIRTainty assay uses native peptides, it can be used to study the effects of previously reported modulators, and to screen for new compounds in a more physiological manner.