| Assay Method Information | |
| | HDAC6 Activity Assay |
| Description: | The enzyme was diluted with 25 mM Tris-Cl, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 0.1 mg/ml BSA, pH 8.0 buffer and its activity was measured by using Substrate 3 (BPS Bioscience; catalog #50037). The assays were carried out in black, low-binding NUNC 96-well plates. Each well contained a volume of 50 μl including buffer, HDAC (50 ng/well), inhibitor, and substrate (20 μM). Prior to adding substrate, the plate was pre-incubated for 5 min. Upon addition of substrate, the plate was incubated at 37 °C for 30 min. At this point, HDAC assay developer (50 μl, BPS Bioscience; catalog #50030) was added to each well and the plate was incubated for 15 min at room temperature. The fluorescence was recorded with a BioTek FLx800 or Synergy H4 microplate reader. |
| Affinity data for this assay | |
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