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| | HDAC Fluorescent Assays |
| Description: | Activity against HDACs 1 to 11 was assessed by using an acetylated 7-amino-4-methylcoumarin (AMC)-labeled peptide substrate. A substrate based on residues 379–382 of p53 (Arg-His-Lys-Lys(Ac)) was used for assaying the activity of HDAC1, HDAC2, HDAC3, HDAC6, HDAC10 and HDAC11. For HDAC8, the diacetylated peptide substrate, based on residues 379–382 of p53 (Arg-His-Lys(Ac)- Lys(Ac)), was used. HDAC4, HDAC5, HDAC7 and HDAC9 assays used the class IIa HDAC–specific fluorogenic substrate (Boc-Lys(trifluoroacetyl)-AMC). The HDACs 1– 11 were purchased from BPS Bioscience Inc. (San Diego, California). All reactions were conducted with a HDAC substrate at 50 μM in assay buffer (50 mM Tris-HCl, pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 1 mg/mL BSA) containing 1% DMSO final concentration. In this assay, a HDAC substrate was deacetylated by the HDAC. Trichostatin A (TSA) was then added to stop the HDAC activity, and the reaction was developed by the addition of trypsin |
| Affinity data for this assay | |
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