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SIRT-Glo™ Assay

A0003

Detects Activity of NAD+-Dependent HDAC Class III Enzymes and SIRTs

  • Simple add-mix-measure protocol
  • Measure deacetylating activities from purified enzyme sources

Size

Catalog number selected: G6450

$ 460.00 Your price: Log In

SIRT-Glo™ Assay
10ml
$ 460.00
Your price: Log In

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
SIRT-Glo Assay chemistry.
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

Nicotinamide

  • 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

Protocols

Specifications

What's in the box?

Item Part # SizeConcentrationAvailable Separately

SIRT-Glo™ Substrate

G644A 1 x 1 vial

Developer Reagent

G653A 1 x 10μl

Nicotinamide

G654A 1 x 30μl1M View Product

SIRT-Glo™ Buffer

G658A 1 x 25ml

SDS

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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

BB

See Protocol for storage recommendations.

Patents and Disclaimers

U.S. Pat. Nos. 6,602,677, 7,241,584, 8,030,017 and 8,822,170 and other patents and patents pending.

U.S. Pat. No. 8,632,992 and other patents pending.

Promega has a non-exclusive, worldwide license to U.S. Pat. Nos. 7,033,778, 7,256,013, Europe Pat. No. 1243568 and Japan Pat. No. 4267043 and related applications to manufacture, have manufactured, use, possess, distribute, market, sell, offer for sale, and import deacetylase activity assay kits and related products for research and laboratory use (including the use on human derived cell lines, but excluding such use on human subjects for diagnostic or therapeutic purposes), product control, process control, product development and process development.

Furthermore, the rights above are extended to purchasers of licensed products from Promega to use, possess, distribute, market and sell such licensed products for the same purposes and for the same scope as granted to Promega.

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