| Assay Method Information | |
| | Human Factor B Assay |
| Description: | CVF-Bb complex, prepared from purified cobra venom factor, (3 nM) is incubated with a compound of the disclosure at various concentrations for 10 minutes at room temperature in PBS (pH 7.4) containing 10 mM MgCl2 and 0.05% (w/v) CHAPS. Human Complement C3 substrate is added to a final concentration of 1 μM. After 1 hour incubation at room temperature, the enzyme reaction is stopped by addition of a cocktail of concentrated pan-protease inhibitors. The product of the reaction, C3a, is quantified by means of an enzyme-linked-immunosorbent assay and/or denaturing gel electrophoresis (SDS-PAGE). IC50 values are calculated from percentage of inhibition of CVF-Bb activity as a function of test compound concentration. |
| Affinity data for this assay | |
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