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The heat change induced by the injection of buffer into the lysozyme solution and that induced by the injection of TbTIM into buffer were subtracted from the experimental (TbTIM into lysozyme); the data are in Fig. 5B.
Background intensity values consisting of only buffer were subtracted from every fluorescence measurement.
The background scattering intensities (of the buffer) were subtracted from the scattering intensities of the protein solutions.
The heats of dilution, determined by injecting the compound into the sample cell containing only buffer, were subtracted from those in compound/DNA titrations to present the corrected binding induced enthalpy changes.
Contributions of the buffer were subtracted from each wavelength scan and the data normalized to molar ellipticity (degrees square centimeter per mole) to account for dilution effects of the initial DNA concentration in the cell.
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The reference titration of small molecules in the buffer was subtracted from the experimental data, and the data were fitted using the Origin 7.0 (OriginLab Corporation) software.
The contribution of buffer was subtracted from experimental spectra.
The spectrum for the buffer was subtracted from the spectrum of the protein.
ThT background signal from buffer was subtracted from corresponding measurements.
All measurements were performed in triplicate, and the fluorescence intensity of the buffer was subtracted from that of the samples.
The heat of dilution of Hrb into buffer was subtracted from the data and titration curves were fitted using ORIGIN (http://www.originlab.com/), yielding values for the stoichiometry N, equilibrium association constant KA (=KD−1), and enthalpy of binding.
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