| Solution Information | help | |
| Enzyme: | Serine/threonine-protein phosphatase | |
| inhibitor: | BDBM64801 | |
| 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 PP5 inhibition potency of compounds identified as active in a previous set of experiments entitled, "Fluorescence-based primary biochemical high throughput screening assay to identify inhibitors of Protein Phosphatase 5 (PP5)" (AID 1987); and inactive 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 biochemical, fluorescence-based assay employs purified PP5c (catalytic domain) and 3-O-methylfluorescein phosphate (OMFP) as the substrate. Cleavage of OMFP by PP5c converts OMFP to the product O-methylfluorescein that fluoresces when measured at an excitation wavelength of 480 nm and an emission wavelength of 540 nm. As designed, compounds that act as PP5 inhibitors will block PP5c catalytic activity, leading to reduced cleavage of OMFP, thereby reducing production | |
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