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The level of binding was quantified using Image J, and results from three separate experiments were fitted in GraghPad Prism (GraphPad Software Inc).
The small endothermic heats of injection in these experiments were fitted to a linear function that was subsequently subtracted from the equivalent integrated heats of the peptide protein binding experiment before fitting.
The degradation experiments were fitted to first order reaction law.
All data in each experiment were fitted simultaneously.
Individual competition binding experiments were analyzed by iterative curve fitting using Prism.
Individual competition binding experiments were analysed by iterative curve fitting using the following equation in Prism v4.03: (2) where Bmin is the non-specific binding, Bmax is the maximum specific binding, [ L] is the concentration of competing ligand and IC50 is the concentration of competing ligand that blocks half of the specific bound radioligand.
Ribosome binding experiments were performed as previously described [24].
All binding experiments were performed twice.
Single-point ligand binding experiments and saturation binding experiments were conducted as described previously.
And the performed of experiment was fit the international, national and institutional animal experiment rules.
Equilibrium binding experiments are conducted to evaluate the binding characteristics of MIPs and templates (BPA and 2,4-D).
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