| Assay Method Information | |
| | Inhibition Assay |
| Description: | The inhibition assays were performed at 25° C. in 50 mM 3,3-dimethylglutarate buffer, pH 7.0, containing 1 mM EDTA with an ionic strength of 0.15M adjusted by NaCl. The salicylic acid based library was screened in a 96-well format at 10 μM compound concentration. The reaction was started by the addition of 50 μl of the enzyme to 150 μl of reaction mixture containing the final Km value of pNPP and various concentrations of the inhibitor in 96-well plate. The reaction was quenched after 10 minutes by the addition of 50 μl of 5N NaOH, and then this reaction mixture was detected for absorbance at 405 nm by a Spectra MAX340 microplate spectrophotometer (Molecular Devices). |
| Affinity data for this assay | |
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