| Assay Method Information | |
| | Fluorescent Assay |
| Description: | hABHD6:Initial Fluorescent Inhibition Assay (3-Point) In each well of a 96-well plate 8 μL of membrane fraction containing full-length hABHD6 (1 μg total protein) was mixed with 168 μL of assay buffer (50 mM Tris-HCl, pH 7.6), and 20 μL of diluted compound (diluted in a dilution buffer consisting of 50% DMSO/50% assay buffer v/v). Each plate was incubated at RT for 15 minutes before adding 4 μL of 1 mM AHMMCE substrate (final concentration of 20 μM AHMMCE and final volume of 200 μL). The reaction was allowed to proceed for 1 hr at RT before the fluorescence was read at λex 360 nm and λem 460 nm and the inhibition calculated. Each experiment can test 8 compounds at three concentrations (usually 10 μM, 1 μM, and 100 nM). |
| Affinity data for this assay | |
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