| Assay Method Information | |
| | FP Assay |
| Description: | FP experiments were carried out at room temperature using no-binding black 384-well microplates. Recombinant full length PARP1 and PARP2 proteins produced in house were diluted to 20 nM and 60 nM, respectively, with assay buffer (50 mM Tris pH 8, 0.001% Triton X100, 10 mM MgCl2, and 150 mM NaCl) and incubated for 4 h with an equivalent volume of the 8 nM fluorescent probe diluted with assay buffer. Fluorescence anisotropy of the probe when bound to the proteins was measured in the presence of test compounds or solvent control and the effect on anisotropy determined. Polarization values were read using an Envision plate reader using excitation and emission wavelengths of 590 and 630 nm, respectively. All FP values are expressed as mP units. |
| Affinity data for this assay | |
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