| Assay Method Information | |
| | Phosphatase Inhibition Assay |
| Description: | Upon the addition of 4 μL of these respective inhibitor stock solutions to the assay buffer, the resulting range in assay inhibitor concentrations were 0.1 μM to1,000 μM for the various 1-amino-5-naphthol-7-sulfonic acid sources, 0.1 μM to 300 μM for RR601 and 0.1 μM to 300 μM for NSC-87877. Appropriate vehicles were added to the blank and negative control wells. A minimum of three replicate wells was run for each condition and at each inhibitor concentration. The replicate wellswere averaged to give a single data point. The reaction was initiated upon addition of 4 μL of a 29 μM enzyme stock which was dispensed to each of the wells except the blanks utilizing a single channel pipette. The resulting DUSP5 PD(WT) concentration in the assay buffer was 0.58 μM. The plate was returned to the plate reader, shaken, and absorbance recorded at 25 °C every thirty seconds over a ten to sixty mins time course. In selected experiments the plate was returned to the plate reader following a 10 mins recording for one or two additional 10 mins absorbance recordings. |
| Affinity data for this assay | |
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