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Assay Method Information

Assay Name:  CA Activity Assay
Description:  Reactions were measured using 0.2 mM phenol red (Absmax at 557 nm) as the indicator, in 10 mM HEPES, 0.1 M Na2SO4, pH 7.5, for a period of 10-100 s. To determine the kinetic parameters and inhibition constants, the CO2 concentration ranged from 1.7 to 17 mM. For the inhibitor assay, at least six traces of the initial reaction were used for determining the initial velocity, which were subtracted from the uncatalysed reaction. A stock solution of 1 mM acetazolamide in 10-20% (v/v) dimethyl sulphoxide (DMSO) was used to prepare dilutions up to 0.01 nM. Inhibitor and enzyme solutions were preincubated prior to inhibition measurements for 15 min at 25C to form the enzyme-inhibitor complex.
Affinity data for this assay
 

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Last update November 1, 2007
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