| Assay Method Information | |
| | Biochemical (FP) Assay |
| Description: | Compounds were dissolved in DMSO an Echo550 liquid handler was utilized to make serial dilations in the desired concentration range in Optiplate-384F plates. 100% DMSO was used for the high (with protein) and low (without protein) control samples. 20 nL of compound or DMSO alone was added to individual assay plate wells. PARP1 and PARP2 protein were expressed, purified, and diluted in assay buffer containing 50 mM Tris pH 8.0, 0.0010% Triton X-100, 10 mM MgCI2, 150 mM NaCl to a final concentration of 20 nM. The PARPi-FL was then added at a final concentration of 3 nM. The assay plate is centrifuged at 1000 rpm for 1 min and incubated for 4 h at room temperature. The fluorescent polarization is read using an Envision plate reader using the following settings:Excitation filter—FITC FP 480-Ex Slot 3Emission filter FITC FP P-pol 535-Em Slot 42nd Emission filter—FITC FP S-pol 535-Em Slot3Mirror module—FITC FP Dual Enh-Slot 1 |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |