HDAC-Glo 2 Assay
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 from cells, extracts or purified enzyme sources. 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). The signal from the assay reaction can be measured within 15-45 minutes after reagent addition with no sample manipulation. The HDAC-mediated luminescent signal is persistent, with a half-life of greater than 2 hours, allowing batch processing of multiwell plates.