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SIRT6-MB was incubated with 50 µL of HDAC assay buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM DTT and 0.2 mM NAD+) containing 5 µg of acetyl-histone H3(K9) peptide (amino acid residues 1–21; H3K9Ac) and 1% fenugreek seed extract for 4h at 37°C.