To determine the SIRT1 activity, we used a SensoLyte® 520 fluorimetric SIRT1 activity assay kit (AnaSpec, Fremont, CA, USA), and the SIRT1 activity was measured with the SensoLyte® 520 fluorimetric SIRT1 activity assay kit. In accordance with the manufacturer’s protocol, different concentrations of the drug and vehicle were incubated with SIRT1 enzymes at 37 °C in the presence of an SIRT1 fluorimetric VICTOR X2 (Perkin Elmer, Waltham, MA, USA), with excitation at 490 nm and emission at 520 nm. The fluorescence was measured using experimental data and calculated using GraphPad Prism7 (GraphPad Software, San Diego, CA, USA).
Sensolyte 520 fluorimetric sirt1 activity assay kit
Manufactured by AnaSpec
Sourced in United States
The SensoLyte® 520 fluorimetric SIRT1 activity assay kit is a tool for measuring the enzymatic activity of SIRT1, a histone deacetylase enzyme. The kit utilizes a fluorogenic substrate that emits fluorescence upon SIRT1-mediated deacetylation. This allows for the quantitative determination of SIRT1 activity in biological samples.
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2 protocols using sensolyte 520 fluorimetric sirt1 activity assay kit
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Fluorimetric SIRT1 Activity Assay
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Fluorimetric Assay for SIRT1 Activity
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SIRT1 activity was measured with the SensoLyte® 520 fluorimetric SIRT1 activity assay kit (AnaSpec, Fremont, CA) according to the manufacturer's instructions. Nicotinamide was used as a reference compound. Briefly, SIRT1 enzymes were incubated with vehicle or various concentrations of test compound at 37oC in the presence of an SIRT1 fluorimetric substrate. The SIRT1 assay developer, which produces a fluorophore in the reaction mixture, was added, and the fluorescence was measured using VICTOR X2 (Perkin Elmer, Waltham, MA, USA) with excitation at 490 nm and emission at 520 nm. The measured activities were calculated using GraphPad Prism (GraphPad Software, San Diego, CA, USA).
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