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Product Details
| Cat # +Size | K324-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Fluorescence (Ex/Em = 400/505 nm) |
| Applications | Detection of Sirtuin Activity in variety of samples |
| Features & Benefits | • Rapid, and convenient • The assay can detect Sirtuin activity as low as 0.06 µU of recombinant human SIRT6. |
| Kit Components | • Sirtuin Assay Buffer • Homogenization Buffer • 1M DTT • Substrate (in DMSO) • NAD • Positive Control • Trichostatin A (in DMSO) • Developer • AFC Standard (in DMSO) (1 mM) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Sirtuins are a class of proteins that possess either histone deacetylase or mono-ribosyltransferase activity. Sirtuins are localized in the cytoplasm, nucleus, nucleolus as well as mitochondria. They are associated with aging, cellular protection, sugar metabolism and cell cycle regulation. Unlike other known protein deacetylases, which simply hydrolyze acetyl-lysine residues, the sirtuin-mediated deacetylation reaction hydrolyzes acetyl-lysine and NAD. This hydrolysis yields the deacetylated substrate, O-acetyl-ADP-ribose 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 Activity Assay Kit, the acetylated p53-AFC substrate is deacetylated by Sirtuins in the presence of NAD+ to generate the deacetylated p53-AFC substrate, nicotinamide and O-Acetyl-ADP Ribose. Cleavage of the deacetylated p53-AFC substrate by the Developer releases the fluorescent group, which is detected fluorometrically at Ex/Em = 400/505 nm. HDAC’s also deacetylate the acetylated p53-AFC substrate. Trichostatin A is added to the reaction to specifically inhibit HDAC’s in samples. This kit provides a rapid, simple, sensitive, and reliable test to measure Sirtuin Activity in a variety of samples. The limit of quantification of the assay is 0.06 µU of recombinant human SIRT6.
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Can the kit determine SIRT1 activity specifically?
Please note there is no way to distinguish SIRT1 activity from other SIRT family members (mainly SIRT3) using this kit without immunoprecipitation
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
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?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
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.
