| Assay Method Information | |
| | Fluorescence Polarization TTR Tetramer Binding Assay |
| Description: | Compound binding to TTR was assessed in the fluorescence polarization assay. The assay measured competitive displacement of the fluorescent probe, FITC-diclofenac, from TTR isolated from human plasma (Clabiochem-Millipore, cat. No. 52957). FITC-diclofenac was synthesized at LeadGen Labs, LLC. Each well contained 200 nM TTR and 100 nM FITC-diclofenac in the FP buffer (10 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.01% CHAPS, 0.01% Prionex) along with test compounds. Nonspecific binding was determined in the presence of 500 μM unlabeled diclofenac (Sigma-Aldrich). Reactions with test compounds were incubated overnight at 4° C. and FP was measured on SpectramaxM5e plate reader (Molecular Devices). |
| Affinity data for this assay | |
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