"Bottom" expresses the amount of unspecific radioligand binding in presence of a very high concentration of competitor.
We performed a non-linear regression analysis of the decay-corrected CPM values to calculate the inhibition constant value (IC50), the concentration of competitor required to inhibit 50% of radiotracer binding, utilizing the GraphPad Prism software program (GraphPad Software, Inc., CA).
In contrast, the double mutant D90mdxR1M was unable to significantly inhibit complex formation even at the highest concentration of competitor tested.
Direct binding isotherms were fit with the concentration of competitor ligand set to zero.
When concentration of competitor dsDNA reached 60 nM nucleotides (same molar concentration as ssDNA), 29.5% of ssDNA was unwrapped.
When a series of experiments is performed using a differing concentration of competitor, the retention time of the analyte will vary.
The concentration of competitor peptide inducing 50% inhibition of marker peptide binding was determined, and expressed as the 50% inhibitory concentration (IC50 in μM).
It is recommended that the sufficient concentration of competitor is determined experimentally for each fraction of fragments or individual fragment and nuclear extract.
The IC50 values (the concentration of competitor required to inhibit 50% of the radioligand binding) of the peptides were calculated by using Origin 8.0 (MicroCal).
The calculated concentration of the target sequence is equal to the concentration of competitor sequence, determined from the regression line, when the log ratio of the products equals zero.
