In vitro SIRT6 deacetylation assay

It's my honor to receive your email, We used Abcam's SIRT6 Activity Assay Kit (Fluorometric) measures the activity of SIRT6 by the basic principle of changing a SIRT6 reaction into the activity of the peptidase In this kit, fluorophore and quencher are coupled to amino terminal and carboxyl terminal of substrate peptide, respectively, and before reaction of deacetylase, the fluorescence cannot be emitted. However, if SIRT6 performs deacetylation, substrate peptide will become cut by the action of peptidase added simultaneously, quencher will separate from fluorophore, and fluorescence will be emitted. Deacetylase enzyme activity is measured by measuring this fluorescence intensity. using 96wells plate with excitation at 480-500 nm and emission at 520-540 nm. Numerous extraction and purification methods can be used to isolate SIRT6. We purified GST-SIRT6 protein in vitro, and then incubated it with our designed peptide. The fluorescence intensity of SIRT6 substrate reflected the deacetylase activity of SIRT6。The specific time of reaction and incubation conditions are shown in the manual