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Following immunoprecipitation, beads were resuspended in 2x urea loading buffer (20 mM Tris-HCl, pH 6.8, 5% SDS, 8 M urea, 100 mM dithiothreitol, and 0.02% bromophenol blue) at 1:1 volume/volume ratios and incubated at 37C for 10 minutes to elute the immunoprecipitated proteins from the beads

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