| Assay Method Information | |
| | Carbonic Anhydrase II Kinetic Assay |
| Description: | Kinetic studies were performed by using different concentration of inhibitors over different concentrations of substrate (4-NPA) such as 0.175, 0.35, 0.70 and 1.40 mM. The enzyme 0.2 mg/mL concentration for each well was used after dissolving in de-ionized water. HEPES-tris ammonia was used as buffer at pH of 7.4. The change in absorbance was measured by keeping the 96-well flat bottom plate in ELISA reader (Spectra max 384, Molecular Devices USA) after addition of substrate for a period of 30 min at 25 °C with 1 min interval. |
| Affinity data for this assay | |
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