| Assay Method Information | |
| | In Vitro Assessment of Peptide Substrate Cleavage by BoNTA-LC |
| Description: | Reactions between recombinant BoNTA-LC and fluorescent peptide substrate were carried out in 96-well microplates. Reaction progress was measured continuously by increase in fluorescence at Ex =398 nm, Em=485 nm as the cleavage of the substrate relieved the quenching of DACIA fluorescence by the 2,4-dinitrophenyl-lysine. The initial velocity values for enzymatic cleavage of peptide substrate were plotted against inhibitor concentration. The points were fit by non-linear regression analysis using the graphing program Prism (GraphPad). IC50 is the inhibitor concentration that achieves half-maximal enzyme inhibition. |
| Affinity data for this assay | |
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