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In order to confirm the specificity of the anti-EGFR-GNP probe in the detection of EGFR from nuclear extract, an inhibition experiment was conducted by treating 5 µl of the sample with 1 µl of monoclonal anti-EGFR antibody at room temperature for 7 min and 24 min prior to using the sample in the assay.
Small amounts of HAs will immediately affect the quantification of low-abundance proteins, especially when the dilution factor for the sample in the assay is limited, unless HA-reducing buffers were included in the test concept during the development phase of the product.
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Immunogenicity assays were designed to obtain low false-negatives in anticipation of eliminating false-positive results by confirmatory assays with the threshold statistically calculated to provide 1% false-positive samples in the assay.
All 26 species C samples were positive, along with 1 from 63 species A and 1 from 8 species B samples in the assay (sensitivity 100%; 98% specificity).
Use 4 μl of the samples in the assay and follow the manufacturer's protocol.
Recoveries from striatal B spiked samples in the assay were in the range of 97.3 – 125.9%.
This new approach significantly reduced variation among samples and reduced the amount of necessary sample used in the assay.
Assay quality was monitored by the internal positive control sample included in the assay kit.
%B/B0-values in extracts of fruiting bodies from H. erinaceus were near to 100% and were independent from the sample concentration in the assay.
The sample concentrations in the assay mixture were 400, 200, 100, 10 μg/ml for the MeOH extract and 20 μg/ml for the isolated compounds.
The sample concentrations in the assay mixture were 200, 100, 10 μg/ml for the extracts and 20 μg/ml for the isolated compounds.
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