HDAC-Glo™ Class IIa and HDAC-Glo™ 2 Assays

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.

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:
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.