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The values for non-specific binding were subtracted from total binding values (with data expressed in terms of relative fluorescence units, RFU).
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To analyze relative Hsp70 binding to TPR proteins (FLAG-IP), first the Hsp70 signal of the control reaction ( = background Hsp70 binding) was subtracted from the Hsp70 coomassie signals of each TPR protein, and these values were then normalized to the coomassie signals of the precipitated TPR proteins.
The non-specific binding was subtracted from all other values.
Non-specific binding was subtracted from total binding to obtain specific binding.
Background binding was subtracted from the value obtained for binding to the consensus DNA sequence.
Background (mock) binding was subtracted from the signal, which was expressed as a percentage of signals present in untreated cells.
Signals from buffer injection and control surface binding were subtracted in all experiments to account for nonspecific binding.
Before analysis of the data, the heat changes accompanying ATPγS binding to CaMKII alone were subtracted from the individual data of enthalpy change accompanying ATPγS binding to NR2B or NR2A saturated CaMKII.
For the analysis, the sensograms from the blank cell, in addition to the sensograms obtained with the running buffer alone were subtracted from the binding to remove the system noise.
In case of non-specific binding, these non-specific MFI values were subtracted from the antigen-specific results.
BSA was used as the blocking agent in the ELISA, and the ODs given for binding of serum IgG to BSA alone were subtracted from the ODs for all candidate antigens; there was no significant difference in binding to BSA by IgG in CS, S and NS sera.
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