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Sirt1 fluorometric assay kit

Manufactured by Enzo Life Sciences
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Sourced in United States

The SIRT1 fluorometric assay kit is a laboratory tool designed to measure the activity of the SIRT1 enzyme. It provides a quantitative assessment of SIRT1 deacetylase activity in a simple, sensitive, and high-throughput format.

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Close spelling variants of this product

Fluor-de-Lys SIRT1 fluorometric drug discovery assay kit Fluorometric assay kit

2 protocols using sirt1 fluorometric assay kit

1

SIRT1 Deacetylase Activity Assay

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Deacetylase activity was measured using the nuclear lysate prepared based on the nuclear extraction protocol from Pierce Biotechnology (Rockford, IL) (15 (link)). The activity was assessed using a SIRT1 fluorometric assay kit from Enzo Life Sciences (Farmingdale, NY) (16 (link)) as described by the manufacturer. Resveratrol, an activator of SIRT1 (17 (link)), stimulated SIRT1 deacetylase activity by twofold (7.2 vs. 3.4 arbitrary fluorescence units/min/μg protein with and without resveratrol, respectively), which validates the assay.
Gao S., McMillan R.P., Jacas J., Zhu Q., Li X., Kumar G.K., Casals N., Hegardt F.G., Robbins P.D., Lopaschuk G.D., Hulver M.W, & Butler A.A. (2014). Regulation of Substrate Oxidation Preferences in Muscle by the Peptide Hormone Adropin. Diabetes, 63(10), 3242-3252.
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2

Ischemic Heart Tissue Sirtuin Activity Assay

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After ischemic heart tissue was collected and homogenized, proteins were extracted. Tissue Sirtuin activity was detected using a SIRT1 fluorometric assay kit (Enzo, New York, USA), according to manufacturer instruction. Samples with 100 g of total protein were incubated with Fluor de Lys-SIRT1 substrate (100 μM), NAD+ (50 μM) at 37 °C for 60 min, and 50 μl mixture of developer and nicotinamide (2 mM) was added by pipetting. Fluorescence kinetics was monitored for 1 h with measuring at 360 nm (excitation) and 460 nm (emission) at 37 °C every 5 minutes37 (link).
Yang X.Y., He K., Pan C.S., Li Q., Liu Y.Y., Yan L., Wei X.H., Hu B.H., Chang X., Mao X.W., Huang D.D., Wang L.J., Hu S.W., Jiang Y., Wang G.C., Fan J.Y., Fan T.P, & Han J.Y. (2015). 3, 4-dihydroxyl-phenyl lactic acid restores NADH dehydrogenase 1 α subunit 10 to ameliorate cardiac reperfusion injury. Scientific Reports, 5, 10739.
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