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Binding data were normalised to the maximal specific binding within each data set and IC50 values were calculated with a single site binding model.
Scatchard analysis of the saturation data (linear regression with Excel Software) was used to yield the maximal specific binding sites (Bmax; fmol mg−1 protein).
Scatchard analysis of the saturation data (linear regression with Excel Software) was used to yield the maximal specific binding sites (Bmax; fmol mg−1 protein) and the affinity constant (Kd; nmol L−1).
Competitor binding was expressed as a percentage of maximal specific binding.
There was also significant binding to TRAILR2-transfected cells with GpL-TNC-TRAILmutR1, but the maximal specific binding was significantly lower than with GpL-TNC-TRAILmutR2 and GpL-TNC-TRAIL.
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The Kd was calculated as the free concentration of radio-ligand at half-maximal specific binding by fitting data to the Hill equation and by linear Scatchard transformation.
Interestingly, from a pharmacokinetic perspective, this may indicate that there may be room for increasing the therapeutic concentration of erlotinib, as at maximal concentration, the specific binding would be absent.
In the hippocampus of NMDA-lesioned rats, [H]corticosterone specific binding progressively increased reaching a maximal value of 448 ± 247 fmol/mg prot at day 5 to then decrease to sham levels at day 15 (Table 1).
However, non-specific binding residues can help specific binding residues to increase binding specificity as well.
Maximal contrast (signal to noise) appeared already after about 30 min, and specific binding peaked at about 20 min.
Briefly, Bmax and Kd of GLP-1/Fc were calculated by following the Specific-Nonspecifc binding algorithm for one site binding, using the formula: Specific binding = Bmax * [L]/ Kd + [L]), where Bmax is the maximal binding at a given [L], [L] = Ligand concentration.
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