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Calorimetric titration of 53BP1-BRCT into p53-core. The protein concentrations of 53BP1-BRCT and p53-core were 311.4 and 53.2 μM, respectively. The experiment was carried out at 8°C in 50 mM Tris-HCl buffer (pH 7.5), 75 mM NaCl, and 1 mM DTT. (Top) The exothermic heat pulse of each injection. (Bottom) The integrated heat data fit to single-site binding model. This gave n = 0.88 ± 0.01 and Ka = (1.6 ± 0.1) × 105 M−1 and thus a Kd of 6.2 ± 0.5 μM.
