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Assay Method Information |
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| | Kinetic Inhibition Assay |
| Description: | LAP was assayed in 75 mM triethanolamine hydrochloride buffer (pH = 8.4, containing 5 mM MgCl2) at 25°C. The substrate (L-leucine-p-nitroanilide) was dissolved in DMSO and added to the assay buffer followed by the enzyme. The hydrolysis of the substrate was monitored by the change in absorbance measured at 405 nm. All the inhibitor solutions were prepared in the assay buffer. The assay solution, which contained 0.05 mL of the substrate solution (0.2, 0.4, 0.6, 0.8, 1 mM final concentrations), 0.25 mL of an inhibitor solution (concentration dependent on the inhibitor), the enzyme solution (20 μg/ml final concentration) and adjusted to a 1 mL final volume. |
| Affinity data for this assay | |
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