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SIRT2 Inhibitor Screening Assay Kit (Fluorometric)

HTS, includes inhibitor
Catalog #: K322
$505.00

Product Details

Cat # +Size K322-100
Size 100 assays
Detection Method Fluorescence (Ex/Em 395/541 nm)
Species Reactivity Mammalian
Applications Screening and characterization of Sirtuin inhibitors.
Features & Benefits • Simple procedure
• Fast and convenient
• The assay is sensitive, reliable, and high-throughput adaptable.
Kit Components • Assay Buffer
• Substrate
• Cofactor
• Developer
• SIRT2 Enzyme
• Inhibitor Control (Nicotinamide)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Sirtuin or Sir2 proteins are a class of proteins that possess either histone deacetylase or mono-ribosyltransferase activity. Sirtuins have been implicated in influencing aging and regulating transcription, apoptosis and stress resistance, as well as energy efficiency and alertness during low-calorie situations. Unlike other known protein deacetylases, which simply hydrolyze acetyl-lysine residues, the sirtuin-mediated deacetylation reaction couples lysine deacetylation to NAD hydrolysis. This hydrolysis yields O-acetyl-ADP-ribose, the deacetylated substrate and nicotinamide, itself an inhibitor of sirtuin activity. Studies suggest that the human sirtuins may function as intracellular regulatory proteins with mono-ADP-ribosyltransferase activity. In BioVision’s Sirtuin inhibitor screening Kit, Sirtuin deacetylates the substrate and then the developer cleaves the deacetylated substrate to release the fluorescent group, which can be detected fluorometrically at Ex/Em = 395/541 nm. In the presence of SIRT inhibitor, the deacetylation will be impeded which prevents the cleavage of the substrate to release the fluorescent group. The kit provides a rapid, simple, sensitive, and reliable test, which is also suitable for high-throughput screening of SIRT2 inhibitors. Inhibitor control (Nicotinamide) is included to compare the efficacy of the test inhibitors.


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Is the acetylated substrate from the cells?
The substrate is provided as a kit component. For easy understanding of the reaction, we have mentioned that it is an acetylated substrate bound to AMC.
What would be a “proper solvent” for the inhibitor? Will any liquid work? Also, what if the inhibitor is already in solution? I might want to test a cell or tissue extract
The type of solvent would depend on the inhibitor you use. For eg., Sirtinol (Cat # 2062-1,-5) can be dissolved in DMSO, ethanol and dimethyl formamide. The assay buffer in which you dilute it will act as the negative control. You can run the solvent alone as another negative control. Your inhibitor in the solvent will act as your expt samples. If you want to use inhibitors added to cell extracts, the cell extract itself will go in the place of the solvent as the solvent control.
Does the Inhibitor Control (Nicotinamide) already contain Assay Buffer? If not, it should be diluted with the same amount of Assay Buffer as the test samples
You can use the nicotinamide control as is. It does not need to be further diluted.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.