Confidence intervals for allosteric model parameters in a single experiment (R3). (A) χ² values plotted against the variable of interest (x) are obtained by fitting the remaining variables to the decimated data set while keeping x fixed The critical χ² value (horizontal orange line) for 95% confidence intervals was obtained from the F distribution: χ²_crit = χ²_min {1 + [m/(n − m)]F(0.05,m,n − m)}, where χ²_min is obtained from fitting all variables, n is the number of data points, and m is the number of adjustable parameters (Kemmer and Keller, 2010). In the particular case of the allosteric model shown here, m = 4 (Δq_L, V_L, W_D, Δq_D). The variables that determine the steep intermediate portion of the Hill plot (Δq_J = 1.67 e_o; V_J = 61.9 mV) were kept constant. The mean and lower and upper confidence limits are indicated by the vertical gray lines. (B) Distributions of parameter values from 200 bootstrap samples. Mean values μ and 95% confidence intervals are indicated by vertical black lines. Confidence intervals were determined from μ ± 1.96σ, where σ is the sample standard deviation.