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| | BChE Inhibition Assay |
| Description: | BChE inhibitions by the carbamate inhibitors were assayed by the Ellman method [Ellman et al., Biochem. Pharm., 7:88-95]. BChE-catalyzed hydrolysis of BTCh in the presence of the carbamate inhibitors and DTNB was followed continuously at 410 nm on a UV-visible spectrometer. The temperature was maintained at 25.0°C by a refrigerated circulating water bath. All inhibition reactions were performed in sodium phosphate buffer (1 mL, 0.1 M, pH 7.0) containing NaCl (0.1 M), acetonitrile (2% by volume), triton X-100 (0.5% by weight), substrate (ATCh for AChE or BTCh for BChE) (50 μM), DTNB, and varying concentrations of inhibitor. Requisite volumes of stock solution of substrateand inhibitors in acetonitrile were injected into the reaction buffer via a pipet. BChE was dissolved in sodium phosphate buffer (0.1 M, pH 7.0). |
| Affinity data for this assay | |
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