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Assay Method Information

Assay Name:  CA Activity Assay
Description:  Carbonic anhydrase activity was assayed by following the change in absorbance at 348 nm of 4-nitrophenylacetate (NPA) to 4-nitrophenylate ion over a period of 3 min at 25°C using a spectrophotometer (Shimadzu UV-VIS) according to the method described by Verpoorte et al. [Verpoorte et al., J. Biol. Chem., 242:4221-4229]. The enzymatic reaction, in a total volume of 3.0 mL, contained 1.4 mL 0.05M Tris-SO4 buffer (pH 7.4), 1mL 3 mM 4-nitrophenylacetate, 0.5 mL H2O and 0.1mL enzyme solution. A reference measurement was obtained by preparing the same cuvette without enzyme solution. The inhibitory effects of the sulphonamide derivatives were examined. All compounds were tested in triplicate at each concentration used. Different concentrations of the compounds were used.
Affinity data for this assay
 

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Last update November 1, 2007
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