There aren’t too many things that you can be sure about these days, but if you’re studying class III HDACs, aka sirtuins, then getting a little SIRTainty™ is now as easy as ordering a kit. The SIRTainty™ Class III HDAC Assay is a new assay, with novel patent pending technology, designed to measure sirtuin activity easily, reliably and with unmatched sensitivity.
What’s a Fluoro for, Anyway?
Unlike other sirtuin assays that only work with only a single, fluorescently tagged substrate, the SIRTainty assay separates the readout from the substrate, instead using nicotinamidase to measure nicotinamide generated during sirtuin-mediated substrate deacetylation. This secondary reaction delivers a direct readout of class III HDAC enzyme activity.
Millipore’s clever approach makes the use a wide variety of unlabeled acetylated peptide substrates possible and provides labs the flexibility to analyze multiple sirtuin isoforms, run inhibitor/activator assays, or even check out enzyme mechanisms. In addition to greater flexibility, detection artifacts from compounds like resveratrol are avoided by eliminating a bulky fluorophore from the substrate.
This winding road leads straight to SIRTanity™
SIRTainty™ Goes User-friendly
The assay includes all the key reagents and components need to run the assay, and comes in a handy 96-well format. This assay is simple to perform and needs no washing, requires minimal hands on time and gives you results in almost no time. To run the SIRTainty™ assay, you just need to mix sirtuin enzyme, NAD+, acetylated peptide substrate, your favorite test compound, and a nicotinamidase enzyme together and let sit for 30 minutes. When the time is up, add in developer reagent and read the signal on a fluorescent plate reader.
Be Certain About Your SIRTs
SIRTainty™ assays were most rigorously tested on sirtuins 1-3, but it can also pick up activity from any of the sirtuin variants 1-7 without a problem. That SIRT specificity is also paired up with more sensitivity than a Notebook fan club. SIRTainty™ sports a limit of detection as low as 0.16U using purified sirtuin – almost five times better than some other assays using labeled substrates.
If you’re sure that you need to be certain about your SIRTs, the EMD Millipore site has all the SIRTainty™ Class III HDAC Assays you need.