| Assay Method Information | |
| | Enzyme Assay |
| Description: | Human recombinant angiotensin-converting enzyme (hrACE) was obtained from R&D Systems (Minneapolis, Minn.). Its enzyme activity was determined using the internally quenched fluorescent substrate ES005: (7-methoxycoumarin-4-yl)acetyl-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-(2,4-dinitrophenyl)-Lys (R&D Systems, Minneapolis, Minn.). The assay buffer was 0.1 M HEPES, pH 7.4, and the temperature was 30° C. The substrate concentration was below Km (≦40 μM). Enzyme rates were determined in an Fmax fluorescence plate reader (Molecular Devices, Sunnyvale, Calif.) using an excitation wavelength of 320 nm and an emission wavelength of 405 nm. |
| Affinity data for this assay | |
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