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, 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.|
Provide Relevant Insight into Compound Effects in Biological Setting: Make better decisions about your compound library early in drug screening. Panel of Screening Tools Allows Comprehensive Screening of HDAC Activity: Easy detection of Class IIa or Isozyme 2 in the same, convenient platform. Highly Sensitive: Feel confident because you can see more. Obtain a dynamic range 10- to 100-fold higher than comparable fluorescence methods. Flexible Format: Determine inhibitor performance in both biochemical and predictive cell-based formats using viable cells or in vitro with cell extracts or purified recombinant enzymes. Simple Measurement of Deacetylating Activities: Easy implementation from benchtop to screening with a single-reagent-addition, homogeneous, add-mix-measure protocol. Fast Data Acquisition in as Little as 15 Minutes: Achieve maximum signal in as little as 15 minutes with persistent glow-type steady-state signal, making the protocol amenable to automation in high-throughput formats and compatible with luminometers without injectors. Robust Detection: Minimize assay interference often encountered with fluorescent assays with robust, bioluminescence-based detection. This technology also allows you to multiplex with cell-health assays, offering more biologically relevant data within a predictive, cell-based context.