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| | PVdQ Inhibition Assay |
| Description: | Briefly, stock solutions of the colorimetric PvdQ substrate (4-nitrophenyl dodecanoate) were prepared in methanol and stock solutions of each n-alkylboronic acid inhibitor in DMSO. Continuous spectrophotometric kinetic assays were used to determine initial hydrolysis rates by mixing substrate (5 μM), and n-alkylboronic acid inhibitor (ranging from 0.002−60 mM depending on the compound, typically with three concentrations above and below each IC50 value) in Na2HPO4 buffer (40 mM) at pH 8.0 and 25 °C, each reaction being made to a final 20% methanol cosolvent10 concentration in a disposable 1 mL polystyrene cuvette). Each reaction was initiated by addition of purified PvdQ (10 nM) and product formation was monitored by measuring the increase in absorbance at 402 nm continuously. |
| Affinity data for this assay | |
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