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| | Bla2 Activity Assay |
| Description: | Bla2, in 50 mM MOPS and 30% (v/v) glycerol, was assayed for substrate activity by adding enzyme at a final concentration of 0.13 μg/mL to solutions ranging from 10-70 μM nitrocefin in 50 mM MOPS (pH 7.0) to a final volume of 1 mL in acrylic cuvettes at 25°C. To assess the ability of thiomaltol to inhibit the activity of Bla2, concentrations of thiomaltol ranging from 0-750 μM were incubated with enzyme for various time periods at the final reaction volume less the volume of nitrocefin necessary to initiate the reaction. Each mixture was incubated anywhere from 0-60 min in 10 min intervals before initiating reactions with nitrocefin (final concentration 36 μM). Reactions were monitored by observing the increase in absorbance at 485 nm and were performed in triplicate. |
| Affinity data for this assay | |
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