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SIRT-Glo Assay

by Promega
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  • Overview
  • Protocols
  • Specifications
  • Resources
  • Predictive, Flexible Assay for HDAC Enzymes and SIRTs

    The SIRT-Glo™ Assay is a single-reagent-addition, homogeneous, luminescent assay that measures the relative activity of the NAD+-dependent histone deacetylase (HDAC) class III enzymes (sirtuins; SIRTs) from purified enzyme sources. The assay uses an acetylated, luminogenic peptide substrate that can be deacetylated by SIRT 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 SIRT-mediated luminescent signal is persistent with a half-life of greater than 3 hours, allowing batch processing of multiwell plates. The SIRT-Glo™ Assay is broadly useful for NAD+-dependent Sirtuin enzymes.

    • Use a single-reagent-addition, homogeneous, add-mix-measure protocol for easy implementation from benchtop to screening.
    • Achieve 10- to 100-fold higher sensitivity than comparable fluorescence methods.
    • Measure maximum signal in as little as 10–15 minutes with persistent glow-type steady-state signal.
    SIRT-Glo™ Assay Chemistry
  • Protocols

    Complete Protocol

    Download PDF

    SIRT-Glo™ Assay System Technical Manual

    PDF (765 KB)

  • 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

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