| Assay Method Information | |
| | Fluorimetric Assay |
| Description: | Inhibitory constants (Ki) and IC50 values were determined under saturating substrate conditions by using a fluorimetric assay (components were purchased separately). This assay system allows detection of a fluorescent signal upon SIRT6 deacetylation of the fluorescent substrate peptide (AMC-p53) with amino acids 379-382 (Arg-His-Lys-LysAc). Sirtuin deacetylation of the substrate renders the fluorophore-substrate bond susceptible to treatment with lysyl endopeptidase. Trypsin cleavage releases the fluorophore, resulting in an increase in fluorescence. Fluorescence intensity was measured on a Synergy HT Multi-Mode microplate fluorescence reader. |
| Affinity data for this assay | |
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