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Assay Method Information |
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| | MetAP Inhibition Assay |
| Description: | The MetAP reaction with Co2+ as a cofactor is coupled to a prolyl aminopeptidase (ProAP) using Met-Pro-p-nitroanilide as substrate. MetAP-catalyzed cleavage of the N-terminal methionine produces prolyl-p-nitroanilide, which is rapidly hydrolyzed by ProAP to release a chromogenic product. This allows the MetAP reaction to be continuously monitored at 405 nm on a UVVIS spectrophotometer. The initial rate was calculated from the early part of the reaction progress curve (<5 min). Activity assays were performed in triplicate using eight to 16 concentrations of substrate or inhibitor. |
| Affinity data for this assay | |
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