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SIRT-Glo™ Assay

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

Number of Assays Per Plate

Product Cat.# # Assays Volume/Assay Plate Size
SIRT-Glo™ Assay G6450 100 100µl 96-well
400 25µl 384-well


Available Separately

Nicotinamide G6540 100 100µl 96-well
400 25µl 384-well
HeLa Nuclear Extract G6570 100 100µl 96-well
400 25µl 384-well


  • Available separately
  • Known inhibitor of SIRTs and used as a positive control inhibitor
  • Supplied at a concentration of 1M in SIRT-Glo™ Buffer

HeLa Nuclear Extract

  • Available separately
  • May be used as an assay chemistry control
  • Supplied at a concentration of 5mg/ml


What's in the box?

Item Part # Size Concentration

SIRT-Glo™ Substrate

G644A 1 × 1 vial

Developer Reagent

G653A 1 × 10μl


G654A 1 × 30μl 1M

SIRT-Glo™ Buffer

G658A 1 × 25ml


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Certificate of Analysis

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Use Restrictions

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

Storage Conditions


See Protocol for storage recommendations.

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