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In vitro SIRT1 deacetylase activity assay

ZY Zhi-qiang Yao
XZ Xin Zhang
YZ Yongqi Zhen
XH Xu-Ying He
SZ Shuangmei Zhao
XL Xi-Feng Li
BY Bo Yang
FG Feng Gao
FG Fu-You Guo
LF Leilei Fu
XL Xian-Zhi Liu
CD Chuan-Zhi Duan
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The in vitro SIRT1 deacetylase activity was determined using a SIRT1 Activity Assay Kit (Abcam, ab156065) following the manufacturer’s protocol42. In brief, Fluoro-Substrate Peptide, Fluoro-Deacetylated Peptide and NAD were successively added to SIRT1 assay buffer followed by incubation with the tested compounds, as well as the developer buffer and recombinant SIRT1 (5 mg) at 37 °C for 60 min. After excitation at 350 nm, emitted light was detected at 460 nm using microtiter plate fluorometer. The fluorescence intensity of the assay buffer was subtracted from each experimental sample.

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