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. Author manuscript; available in PMC: 2012 Jan 2.
Published in final edited form as: DNA Repair (Amst). 2010 Oct 16;10(1):56–64. doi: 10.1016/j.dnarep.2010.09.007

Figure 4.

Figure 4

Active site titration and determination of KdDNA for DNA Pol κ in the presence of MgCl2 or MnCl2. DNA pol κ (100 nM) was incubated with increasing concentrations of template-primer DNA (6–300 nM) and 500 μM dCTP at different reaction times in the presence of MgCl2 (A–B) or MnCl2 (C–D). The data were fit to the burst equation (A and C). The burst amplitudes ([E:P/T]) were plotted versus template-primer concentrations ([P/T]) (B and D) and fit to the quadratic equation (y = (0.5(Kd+Et+Dt))−((0.25(Kd+Et+Dt)2)−(EtDt))1/2). The active enzyme concentration was determined to be approximately 116 ± 7 nM.