2012 Promega catalogue
2012 Promega catalogue
2012 Promega catalogue
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Cell Signaling<br />
SIRT-Glo Assays and Screening Systems<br />
Product Size Cat.# Price ($)<br />
SIRT-Glo Assay 10 ml G6450 578.00<br />
5 × 10 ml G6451 2137.00<br />
100 ml G6452 2781.00<br />
SIRT-Glo Screening System 10 ml G6470 615.00<br />
5 × 10 ml G6471 2280.00<br />
Product Size Conc. Cat.# Price ($)<br />
SIRT-Glo Control Substrate 35 µl G6460 352.00<br />
HeLa Nuclear Extract 10 µl 5 mg/ml G6570 165.00<br />
Nicotinamide 30 µl 1 M G6540 112.00<br />
For Research Use Only. Not for Use in Diagnostic Procedures.<br />
Description: The SIRT-Glo Assays and Screening Systems are singlereagent-addition,<br />
homogeneous, luminescent assays that measure the relative<br />
activity of the NAD + -dependent histone deacetylase (HDAC) class III enzymes<br />
(sirtuins; SIRTs) from purified enzyme sources. The assays use an acetylated,<br />
luminogenic peptide substrate that can be deacetylated by SIRT activities.<br />
Deacetylation of the peptide aminoluciferin substrate is measured using a<br />
coupled enzymatic system in which a protease in the Developer Reagent<br />
cleaves the deacetylated peptide from the aminoluciferin substrate, releasing<br />
aminoluciferin, which is quantified in a reaction using Ultra-Glo recombinant<br />
firefly luciferase. The assay reaction is typically complete within 15–45<br />
minutes with no sample manipulation. The SIRT-mediated luminescent signal is<br />
persistent with a half-life of greater than 3 hours, allowing batch processing of<br />
multiwell plates. The SIRT-Glo Assay is broadly useful for NAD + -dependent<br />
Sirtuin enzymes.<br />
Nicotinamide, included in the SIRT-Glo Screening Systems or available<br />
separately, is a known inhibitor of SIRTs and used as a positive control inhibitor.<br />
Nicotinamide is supplied at a concentration of 1M in SIRT-Glo Buffer.<br />
The HeLa Nuclear Extract, included in the SIRT-Glo Screening Systems or<br />
available separately, may be used as an assay chemistry control. HeLa Nuclear<br />
Extract is supplied at a concentration of 5mg/ml.<br />
The SIRT-Glo Control Substrate, only available separately, is a non-acetylated<br />
form of the SIRT-Glo Substrate with the same amino acid sequence and can<br />
be used with the SIRT-Glo Assays and Screening Systems to confirm true<br />
SIRT inhibition in secondary screens. The Control Substrate is supplied at a<br />
concentration of 10mM and is sufficient for 480 assays in 96-well plate format<br />
when combined with the SIRT-Glo Reagent prepared with components in the<br />
SIRT-Glo Assays or Screening Systems.<br />
For complete and up-to-date product information visit: www.promega.com/catalog<br />
Features:<br />
• Simple Measurement of Deacetylating Activities: Use a singlereagent-addition,<br />
homogeneous, add-mix-measure protocol for easy<br />
implementation from benchtop to screening.<br />
• Highly Sensitive: 10- to 100-fold higher sensitivity than comparable<br />
fluorescence methods.<br />
• Fast Data Acquisition: Measure maximum signal in as little as<br />
10-15 minutes with persistent glow-type steady-state signal.<br />
Storage Conditions: Store the SIRT-Glo Assay components and SIRT-Glo<br />
Control Substrate at –20°C. Store HeLa Nuclear Extract at –70°C.<br />
Protocol Part#<br />
SIRT-Glo Assay and Screening System Technical Manual TM336<br />
Signal-to-Noise Ratio<br />
10,000<br />
1,000<br />
100<br />
10<br />
SIRT1<br />
SIRT2<br />
SIRT3<br />
SIRT4<br />
SIRT5<br />
SIRT6<br />
1<br />
1 10100 1,000<br />
Recombinant SIRT Enzyme (ng/ml)<br />
Broad linearity and isoenzyme utility.<br />
8910MA<br />
53<br />
3<br />
Cell Signaling<br />
Section<br />
Contents<br />
Table of<br />
Contents