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hSTINGgroup1234 showed a comparable exothermic binding curve and binding affinity (KD: 0.69 μM) to mSTING (KD: 0.49 μM) (Gao et al., 2013b).
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By determining the percentage of the TfR-containing complexes formed at varying mixing ratios, it is possible to obtain a binding curve and thus the binding constant.
This may be compared to a 60-fold increase demonstrated for 1. Attempts to fit these small spectral changes to binding models failed to produce reliable saturation binding curves and thus a DNA binding constant could not be derived for 2 using this latter technique.
(A ) Fluorescence binding curve showing binding of TagRFP-labelled Coh to wild-type and mutant Doc nonspecifically immobilized in a 96-well plate.
Binding curves obtained when Cy5 was positioned at either the 5′ or 3′ terminus closely resemble single site binding curves, whereas binding of XPD to centrally positioned Cy5-labelled oligonucleotides appeared more complex.
The binding mechanism, binding constants, binding sites and binding distance were obtained.
For competition binding, all data points on each binding curve were performed in triplicate, and each 96-well plate also contained 6 determinations of total and nonspecific binding.
BPA half-lives in the exposure tanks were calculated using a one-phase exponential decay model, IC50 (median inhibitory concentration) values with a homologous competitive binding curve for one class of binding sites, and EC10 and EC50 values with a Weibull distribution (Prism 4.02).
Select "One site binding" under the "Nonlinear Regression dialog" box to analyze the data and produce a binding curve.
Preliminary analysis of the binding curves indicated that the binding rates were dominated by mass transfer, and global fitting of the binding curve returned values with low significances (see below).
With serial dilution, the binding curve reflecting the binding ability of mAb products is obtained, and the EC50 value can be derived by four-parameter logistic fit (Fig. 2C) with the use of GraphPad Prism 5.0 (GraphPad Software, San Diego, CA).
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