Fluor-de-Lys^® Green SIRT5 fluorometric green drug discovery assay kit by Enzo Life Sciences

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, N^ε-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.