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The mouse β-actin and β-2 microglobulin values were employed for normalisation of the Q-PCR data.
A mouse monoclonal antibody to cytokeratin 19 (CK19) (0.5 μg ml−1, cloneA53-B/A2.26; Merck ChemicaLtd, Nottinghamham, UK) was employed for normalisation of epithelial protein loading.
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The normalisation factors previously obtained by isotopic labelling were employed for calculation of relative S100 protein expression, thereby keeping maximum precision in this preceding step and allowing the analysis of S100 proteins not detected in the SILAC spike-in standard or not added as heavy peptides before tryptic digestion.
Therefore, we employed normalisation in order to increase the number of unique transcripts discovered [ 37].
For normalisation, the gene Ubiquitin C (UBC) was employed.
For normalisation of gene expression data, GAPDH was employed but the use of alternative house-keeping genes such as 18s ribosomal RNA, Cyclophilin G, and β-actin did not change the results of our analysis (Supplementary Fig. 4).
EGM-2 was employed as a positive control, whereas non-treatment controls were performed for normalisation and comparison.
Our findings identified 2 reference markers that exhibited stable expression across human liver tissues with different conditions thus should be regarded as reliable reference moieties for normalisation of gene and protein expression in clinical research employing human hepatic tissues.
Japan's desire for "normalisation" aims only to build on those benign influences.
Many of those who call for "normalisation" argue that Germany must assert its interests.
For normalisation, two reference genes were used and normalisation factor was calculated by geNorm software (Vandesompele et al. 2002).
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