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The murine plasma samples were subjected to FPLC fractionation analysis with two tandem superose 6 columns (GE Healthcare, Vienna, Austria), as described previously [20].
As a surrogate, we used FPLC lipid profiles from pooled murine plasma samples to differentiate between the high-density and low-density lipoprotein cholesterol fractions in the different treatment arms.
Murine plasma samples from treated and untreated xenografts were analysed by surface-enhanced laser desorption/ionisation time-of-flight mass spectroscopy, and panels of peaks were found using class prediction models that distinguished treatment groups.
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Profiles of the murine plasma sample proteins obtained using SELDI TOF MS on the two (CM10 and Q10) chip surfaces allowed detection of 189 peaks that were present in most samples.
Murine or human plasma samples were analyzed by Millipore Corp. (Billerica, MA, USA) for cytokine/chemokine/growth factor measurements using the Luminex xMAP technology.
In another enzyme protection assay, before the addition of murine RNase inhibitor, plasma samples were incubated with 10 μg/mL RNase A under various conditions, namely, at 37°C for 15 min, at room temperature for 30 min, or at 4°C overnight, followed by exosome isolation and RNA extraction.
The Biocrates Bile Acids Kit utilizes Acquity UPLC coupled to a Xevo TQ-S triple quadrupole mass spectrometry by Waters Corporation to perform quantitative multiplexed analysis of up to 16 human or 19 murine bile acids in plasma samples.
These data support the profile seen in human plasma samples from murine typhus patients in this study, which is also in line with the evidence for the endothelial tropism of R. typhi derived from in vitro experiments, mouse studies, and human post-mortem studies [ 7, 30].
The method of analysis was validated for murine plasma, brain, liver, and kidney samples.
Three of the eight proteins (serum amyloid A (SAA), transthyretin and apolipoprotein A-1) identified in the murine plasma were confirmed in the human samples, with the identification of one additional protein (retinol-binding protein 4) in the human samples only.
Plasma samples consisted of 250 µl of thawed murine plasma and underwent SPE only.
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