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| | Fluorescence-Based CA II Binding Assay |
| Description: | The assay was performed according to a previously reported protocol.[Banerjee et al., Biochemistry, 44:3673-3682] DNSA stock solution was prepared at 1 mm in DMSO/water (1:1). CA II (1 mm) was added to the test probe (over a 0.1-100 mm concentration range) in HEPES buffer [HEPES (25 mm) with acetonitrile (10 %), pH 7.0] and mixed well. DNSA (0.1 mm) was added, and the resulting solution was incubated at RT for 10 min and then transferred into a black-wall 96-well plate (final volume 100 mL/well). The fluorescence was measured with a Spectra Max 250 plate reader (Molecular Devices, Sunnyvale, CA, USA) at an excitation of 285 nm and emission of 465 nm at 25 °C. |
| Affinity data for this assay | |
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