| Assay Method Information | |
| | Fluorescent Polarization (FP) ASGPR Binding Assay |
| Description: | The assay was performed in 384-well microplates (GREINER bio-one, 781076). Membrane prepared from Wistar rat liver was used as a source of ASGPR and Cy5 fluorophore-labeled trimeric GauNAc-tool compound was used as a tracer.In the FP binding experiments, 15 μltest sample was mixed with 15 μl of 65 ag liver membrane. After 10 minutes incubation at room temperature, 10 μl of the Cy5-labeled trimeric GalNAc-tool compound (final concentration: 1 nM) were added. The FP signals were recorded after 30 minutes incubation at room temperature at Ex. 612 nm/Em. 670 with PheraStar FXS (BMG Biotech). The optimal FP buffer was 50 mM Tris, pH 7.4; 3 mM CaCl2; 0.04% Triton X100; 0,05% BSA. |
| Affinity data for this assay | |
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