Fig. S3.

Further purification of disassembly factor by gel filtration chromatography. Partially purified disassembly factor from pooled fractions 21 and 22 of anion exchange chromatography (2 mg of protein) were applied to a Superdex 200 Increase 10/300 GL column (GE Healthcare), equilibrated with 50 mM Tris⋅HCl (pH 7.2), 150 mM NaCl, 1 mM DTT, 10% (vol/vol) glycerol, and 0.1 mg/mL BSA. Fractions of 500 μL were collected at a flow rate of 0.5 mL/min. Samples of 3 µL of the indicated fractions were assayed for Cdc20 release from anti–BubR1-bound BC-1 as described in Materials and Methods. Arrows at the Top indicate the elution position of marker proteins (in kilodaltons).