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. 2013 May;11(4):253–268. doi: 10.1089/adt.2012.495

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Copyright 2013, Mary Ann Liebert, Inc.

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Fig. 4. — Optimal bead concentrations for AF4-AF9 binding assays. (A) To determine optimal bead concentrations, biotin-AF4 27-mer and AF9-FLAG (2.4 nM each) were preincubated for 90 min, followed by 60-min incubation with varying concentrations of AlphaScreen beads. Controls (●) consisted of biotin-AF4 27-mer and AF9-FLAG; background (■) also included 300 nM nonbiotinylated AF4 27-mer. Values=mean±SD; n=3. (B) AF4-AF9 AlphaScreen control signals for 8 and 20 μg/mL bead concentrations in a total 12-μL assay volume. Biotin-AF4 27-mer and AF9-FLAG (2.4 nM each) were preincubated for 90 min, followed by 60-min incubation with AlphaScreen beads. Controls are shown in gray and background shown in black. Values=mean±SD; n=24.