In vitro GST pulldown assays were performed in 50 µl volumes in binding buffer (25 mM Hepes pH 7.4, 100 mM NaCl, 0.5 mM MgCl2, 0.01% NP40), ∼10 µg GST protein fusion bound to a glutathione Sepharose support, 100 ng human NURF complex. Proteins were allowed to bind for 1 hour at 4°C with occasional mixing. The beads were washed with 500 µl binding buffer three times at 4°C and re-suspended in a final volume of 30 µl.
