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Chemokine proteins were quantified in reference to serial dilutions of recombinant standards falling within the linear part of the standard curve for each specific chemokine sample measured.
Quantitative assay results falling within the linear range of the assays were reported in copies/mL.
TSLP protein was quantified with reference to serial dilutions of the recombinant standards, falling within the linear part of the standard curve for each specific TSLP value measured.
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Most clinical specimens tested (40/41) fell within the linear range of the assay.
Specimens were chosen on the basis of having fallen within the linear range of the first assay.
The number of cycles used was optimized for each gene to fall within the linear range of PCR amplification.
The amount of protein loaded was adjusted for each antibody so that signal intensity fell within the linear range of the assay (Supplemental Figure S2).
Multiple serial dilutions of a reaction were fractionated alongside the undiluted reaction, and the substrate band was quantified from those that fell within the linear range of estimation.
Test samples that did not fall within the linear part of the optical density range of the standard were tested at alternate dilutions.
The band intensity in the input lanes and biotin lanes, which fell within the linear range, was quantified to calculate the % of total GluR1 or GluR2 on the surface for each sample.
The spot intensities of the control wells were plotted against the antibody concentration (Ablinear, rabbit) or antiserum dilution (Abcyclic, mouse), and values that fell within the linear regions of the curves were used to normalize for slight differences in the number of bacteria in each spot.
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