CHEMBL5730547

Fluorescence Detection: Enzymatic activity was tested in 96-well or 384-well flat microwell plate by fluorescence detection and taking Ac-Lys-Tyr-Lys(Ac)-AMC as substrate. The tested samples was firstly formulated to be a 10−2 M stock solution in DMSO, subpackaged and storaged at −20° C., and then diluted to required concentration with reaction buffer before use. Substrate Ac-Lys-Tyr-Lys (Ac)-AMC was deacetylated by HDAC1, and the fluorescence signal of the product AMC obtained by enzyme hydrolysis was detected at 355 nm or 460 nm by a fluorescence detector. The initial reaction speed was calculated by detecting the change of fluorescence signal with time.