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A mixture of 50 nM of His-tagged biotin (Met-His-His-His-His-His-His-Ser-Ser-Gly-Val-Asp-Leu-Gly-Thr-Lys-(Biotin)), streptavidin-coated donor beads and Ni-chelate acceptor beads were distributed into 96-well plate. Both of beads bound to the His-tagged biotin. Then 3478 was added in a series of dilution to test the compound’s interference of binding between the streptavidin-coated donor beads and the AlphaScreen Ni-chelate acceptor beads with the His-tagged biotin, by monitoring light signal between 520 and 620 nm.
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