| Assay Method Information | |
| | Inhibition of Histone Deacetylase Enzymatic Activity |
| Description: | Below is a standardized protocol for running HDAC selectivity panel on Caliper LabChip EZ-Reader Instrument.The Caliper HDAC Assay Buffer (acronym HAB, 1 liter) was prepared as follows:Final Components: Concentration: Catalog #s:100 mL 1M KCL 100 mM Sigma #9541-500G 50 mL 1M HEPES, 50 mM Sigma #H3375-1KGpH 7.4 1 mL 10% BSA 0.01% SeraCare #AP-4510-80-100G 20 μL 50% Tween-20 0.001% Zymed #00-3005-20mLThe components were added to 1 liter Milli-Q water and store at 4° C.The substrate (stock conc.) was prepared as follows:Substrate A was prepared as 2 mM in DMSO. Its final concentration in the assay for HDACs 1,2,3,6 is 2 μM.Substrate B was prepared as 2 mM in 100% DMSO. Its final concentration in the assay for HDACs 4,5,7,8,9 is 2 μM.LBH was used as quench inhibitor to stop the reaction at the end point. The instrument buffer was ProfilerPro Separation Buffer (e.g., Caliper #760367). The instrument chip was LabChip EZ Reader II 12-Sipper Off-Chip Mobility Shift Chip (e.g., Caliper #760404). |
| Affinity data for this assay | |
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