SIRT1 Deacetylase Activity Assay

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.

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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.

Publication 2014

SIRT1 fluorometric assay kit

Assay Fluorescence Fluorometric Protein Resveratrol Sirt1

Corresponding Organization : Saint Louis University

Other organizations : Virginia Tech, Universitat Internacional de Catalunya, Centro de Investigación Biomédica en Red, Universitat de Barcelona, University of Alberta

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Variable analysis

independent variables

Resveratrol (an activator of SIRT1)

dependent variables

SIRT1 deacetylase activity

control variables

Nuclear lysate prepared based on the nuclear extraction protocol from Pierce Biotechnology
SIRT1 fluorometric assay kit from Enzo Life Sciences

controls

Positive control: Resveratrol stimulated SIRT1 deacetylase activity by twofold, which validates the assay.
Negative control: Not explicitly mentioned.

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