SIRT1 (Sirtuin1) Fluorogenic Assay Kit
The Fluorogenic SIRT1 Assay Kit is a complete assay system designed to measure Sirtuin 1 (SIRT1) activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent SIRT1 (Sir2) activity measurements. In addition, the kit includes purified SIRT1 enzyme and a SIRT inhibitor, Nicotinamide, for use as a positive and negative control, respectively. The Fluorogenic SIRT1 Assay Kit is based on a unique fluorogenic substrate and developer combination. This assay method eliminates dealing with the radioactivity, extraction, and chromatography aspects of traditional assays. Using this kit, only two simple steps on a microtiter plate are needed to analyze the SIRT1 activity level. First, the HDAC fluorometric substrate (HDAC substrate 1), containing an acetylated lysine side chain, is incubated with purified SIRT1 enzyme. The deacetylation sensitizes the substrate so subsequent treatment with the Lysine Developer produces a fluorophore that can then be measured using a fluorescence reader.
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This product has been cited 14 times.
Catalog Number |
Reagent |
Amount |
Storage |
|
50012 | SIRT1 human recombinant enzyme | 50 µg | -80°C | Avoid freeze/ thaw cycles! |
50032 | Fluorogenic HDAC substrate (5 mM) | 50 µl | -80°C | |
Nicotinamide Adenine Dinucleotide (NAD+) (50 mM) |
50 µl |
-80°C |
||
Nicotinamide (10 mM) | 500 µl | -80°C | ||
50089 | 2x SIRT Developer (contains 2 mM Nicotinamide) |
6 ml |
-80°C |
|
50090 | SIRT assay buffer | 10 ml | -20°C | |
79685 | Black, low binding NUNC black microtiter plate | 1 plate |
Room Temp. |
1. A. Ito et al. (2001) EMBO J. 20 1331.
2. A. Ito et al. (2002) EMBO J. 21 6236.
3. N.A. Barlev et al. (2001) Mol. Cell 8 1243.