Assay Method Information |
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| Inhibition Assay |
Description: | All assays were performed at 100 µL total volume in triplicate and monitored for 60 minutes at room temperature in a 96-well plate format in HEPES Buffer (100 mM HEPES, 100 mM NaCl, 0.1% CHAPS, 10% sucrose, pH 7.5) on a BioTek Synergy H1 plate reader. Fluorescence was measured with excitation and emission wavelengths of 380 nm and 460 nm, respectively. Caspase-1 was activated with 100 µM DTT for 30 minutes and held constant at 5 nM for all assays. WEHD-AMC was used as a substrate for all assays at 10 µM unless otherwise specified and reactions were monitored immediately upon addition to caspase-1. |
Affinity data for this assay | |
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