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Assay Method Information |
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| | SpeB and Papain Kinetic Assay |
| Description: | SpeB was incubated at 50 nM in the presence of increasing amounts of inhibitor (3 μM to 200 μM) in a reaction buffer consisting of PBS at pH 7.4, 0.1 mM EDTA, 10 mMDTT, and 0.1% CHAPS and incubated for 30 min at 25 °C. Fifty micromolar Ac-AIK-AMC was subsequently added, and the rate of substrate hydrolysis was measured by increase in fluorescence every 20 s for a 5 min duration in 96-well plates on a PerkinElmer EnVision plate reader. |
| Affinity data for this assay | |
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