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. Author manuscript; available in PMC: 2014 Mar 24.
Published in final edited form as: Biochem J. 2011 Mar 15;434(3):415–426. doi: 10.1042/BJ20101437

Figure 3. Use of a PEG precipitation assay to identify binding of InsP6 and PtdIns(3,4,5)P3 to PBD2.

Figure 3

PBD2 (32 μg) was incubated with 3HInsP6 and increasing amounts of either non-radioactive InsP6 (circles) or C8 -PtdIns(3,4,5)P3 (squares) as described in the Materials and methods section. Bound and free ligand was separated by a PEG precipitation technique (see Materials and methods section). The Kd for InsP6 (0.57 μM) was calculated from the saturation binding plot (inset). The value of Bmax was 0.6 mol/mol of protein.