The HDAC-Glo™ Class IIa and HDAC-Glo™ 2 Assays are single-reagent-addition, homogeneous, luminescent assays that measure the relative activity of histone deacetylase (HDAC) Class IIa and Class I enzyme 2, respectively, in the same, convenient platform using purified enzymes, extracts or cells directly in culture plates. The assays can be used to determine HDAC inhibitor potency, selectively profile HDAC inhibitors, correlate HDAC inhibitor potency with cellular fate in same-well multiplexed viability assays or determine off-target HDAC effects of compounds.
The assays use an acetylated, live-cell-permeant, luminogenic peptide substrate that can be deacetylated by HDAC activities. Deacetylation of the peptide aminoluciferin substrate is measured using a coupled enzymatic system in which a protease in the Developer Reagent cleaves the deacetylated peptide from the aminoluciferin substrate, releasing aminoluciferin, which is quantified in a reaction using Ultra-Glo™ Recombinant Luciferase (firefly). This technology also allows you to multiplex with cell-health assays, offering more biologically relevant data within a predictive, cell-based context.