Recombinant HDAC1 protein gel. HDAC1 protein was run on a 10% SDS-PAGE gel and stained with Coomassie blue. MW: 56 kDa Purity> >65%
Recombinant HDAC1 activity by HTRF Assay. 1 μM H3K9ac (1-21aa.) peptide was incubated with different concentrations of HDAC1 protein in 10 μl reaction system containing 25 mM Tris-HCl pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 0.1 mg/ml BSA for 30 min at room temperature, then 10 μl anti-H3K9me0 antibody and SA-XL665 mixture (1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min at room temperature. All operations and reactions were performed at room temperature. HTRF assay was used for detection.
Recombinant HDAC1 activity by HTRF Assay. 1 μM H3K9ac (1-21aa.) peptide was incubated with different concentrations of HDAC1 protein in 10 μl reaction system containing 25 mM Tris-HCl pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 0.1 mg/ml BSA for 30 min at room temperature, then 10 μl anti-H3K9me0 antibody and SA-XL665 mixture (1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min at room temperature. All operations and reactions were performed at room temperature. HTRF assay was used for detection.