The Fluor de Lys®Green SIRT5 Fluorometric Drug Discovery Kitis a complete assay system designed to measure the lysyl desuccinylase activity of the recombinant human SIRT5 included in the kit. A black 96-well microplate is packaged with the kit, but it should be noted that reagents of the Fluor de Lys®system have also been successfully employed in other formats, including cuvettes and 384-well plates.
The Green SIRT5 Fluorescent Activity Assay is based on the unique Fluor de Lys®-Succinyl GreenSubstrate/Developer combination. The assay procedure has two steps. The Fluor de Lys®-Succinyl GreenSubstrate, which comprises a lysine residue, Nε-succinylated on its side-chains, is first incubated with human recombinant SIRT5 together with the cosubstrate NAD+. Desuccinylation of Fluor de Lys®-Succinyl Green sensitizes it so that, in the second step, treatment with the Fluor de Lys®Developer produces a fluorophore. Use of a succinylated, rather than acetylated substrate with SIRT5 results in readily observed saturation kinetics and a greater than 1000-fold increase in assay sensitivity.
Manufacturer Enzo Life Sciences
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