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Changes in free energy (ΔG) and entropy (ΔS) upon binding were calculated from the KD and ΔH as described in Methods.
The apparent dissociation constants for hematein binding were calculated from secondary plots of the slopes or intercepts obtained from the Linewear-Burk plots.
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Percent of binding was calculated from the resulting number of pixels for each of the investigated factors.
Specific binding was calculated from the difference between the integrated radioactivity signals per area (DLU/mm2) under baseline and blocking conditions, divided by the value under baseline conditions.
Percent of gp120-CD4 binding was calculated from gp120-CD4 complex formation in the absence of any inhibitor.
Absorbance was measured at 415 nm and relative binding was calculated from background-subtracted values.
Specific binding was calculated from the difference between radioactivity levels measured in the absence and presence of unlabelled oestradiol.
As a consequence, the apparent second-order rate (kon) constant for cyanide binding was calculated from the slope of the linear plot of kobs(1) versus cyanide concentration.
The percent E6 E6AP binding was calculated from raw luminescent values for each test compound, and compounds that displayed 3 standard deviation units (∼50% inhibition) from the average of E6 E6AP binding in the presence of DMSO were assigned as active.
Specific binding was calculated from the difference between radioactivity levels measured in the absence and presence of unlabelled oestradiol and the data were analyzed according to the method of Scatchard [ 30].
Δ S° is the standard change in entropy upon binding was calculated from determined equilibrium parameters using the equation: − RTLn KA)=Δ G°=Δ H° − TΔ S°, where R is the universal gas constant (1.9872 cal·mol−1·K−1), T is the temperature in Kelvin, KA is the association constant.
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