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Assay Method Information |
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| | Biolayer Interferometry Binding Assay |
| Description: | For this experiment, we immobilized the biotinylated recombinant Pax2 paired domain, containing amino acids 1−81, onto streptavidin biosensors and measured the binding affinity of EG1 to the immobilized Pax2 paired domain. After biotinylation, the protein was bound to saturation to the biosensor of an Octet RED Multichannel platform (ForteBio). All binding and equilibration was done in PBS with 0.1% DMSO. For binding to the compound, sensors were incubated for 300s in EG1 at the following concentrations: 0.5, 1, 2, 4, 8, 16, 31, 62, 125, and 250 μM. Disassociation was then measured for 350 s in PBS to determine affinity constants. Kd is calculated from Kon and Koff. |
| Affinity data for this assay | |
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