| Solution Information | help | |
| Enzyme: | Trans-activator protein BZLF1 | |
| inhibitor: | BDBM64839 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine the selectivity of compounds identified as active PP1 inhibitors in a previous set of experiments entitled, "Counterscreen for inhibitors of PP5: fluorescence-based biochemical high throughput primary assay to identify inhibitors of Protein Phosphatase 1 (PP1)" (AID 2235). This assay determines the PP5 inhibition potency of tested compounds. This biochemical, fluorescence-based assay employs purified PP5c (catalytic subunit) and 3-O-methylfluorescein phosphate (OMFP) as the substrate. Cleavage of OMFP by PP5c converts OMFP to the fluorescent product 3-O-methylfluorescein. As designed, compounds that act as PP5 inhibitors will block PP5 catalytic activity, thereby decreasing OMFP cleavage and production of fluorescent product, resulting in reduced well fluorescence. Compounds are tested in triplicate in a 10-point 1:3 dilution series starting at a nominal test concentration of 67 micromolar. Protocol Summary: Prior to the sta | |
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