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HDAC-Glo I/II Assay

by Promega
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  • Overview
  • Protocols
  • Specifications
  • Resources
  • Predictive, Flexible Assays for HDAC Activity

    The HDAC-Glo™ I/II Assays and Screening Systems are single-reagent-addition, homogeneous, luminescent assays that measure the relative activity of histone deacetylase (HDAC) class I and II enzymes using purified enzymes, extracts or cells directly in culture plates. The add-mix-measure protocol allows for simple measurement of deacetylating activities and easy implementation from benchtop to screening. The HDAC-Glo™ I/II Assays and Screening Systems 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 firefly luciferase.

    The assay reaction is typically complete within 15–45 minutes with no sample manipulation. The HDAC-mediated luminescent signal is persistent, with a half-life of greater than 3 hours, making the protocol amenable to automation in high-throughput formats and compatible with luminometers without injectors and allowing batch processing of multiwell plates. The HDAC assay is broadly useful for class I and II enzymes. The HDAC-Glo™ I/II Assays are highly sensitive, allowing you to obtain a dynamic range 10- to 100-fold higher than comparable fluorescence methods.

    HDAC-Glo™ I/II Assay Chemistry
  • Protocols

    Complete Protocol

    Download PDF

    HDAC-Glo™ I/II Assay and Screening System Technical Manual

    PDF (1 MB)

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    -30C TO -10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources