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Estimates of the mean differences of molecular masses using Sha31 antibody were −0.66 kDa (p<0.0001), −0.79 kDa (p<0.0001) and −0.36 kDa (p<0.0001) for the bi-, mono- and un-glycosylated bands respectively.
Estimates of the mean differences of molecular masses using Sha31 antibody were respectively +0.32 kDa (p = 0.012), +0.45 kDa (p = 0.014) and +0.65 kDa (p = 0.0005) for the bi-, mono- and un-glycosylated band.
The continuous c(s) analysis method (using a frictional coefficient of 1.9) was used to determine sedimentation coefficients and molecular masses using the SEDFIT software [29].
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To provide an enriched fraction of RFB homogenate for mass spectrometric analysis, the RFB homogenate was separated into 110 fractions by molecular mass using a size fractionation column.
Since the non-uniform fold of rfhSP-D, comprising globular, coil-coiled and extended regions, may lead to an over-estimation of the molecular mass using SEC, we further analysed the protein using ESI-MS.
(B) Determination of molecular mass using gel filtration chromatography.
However, palmitoylation of the protein does not result in a large difference in relative molecular mass using gel electrophoretic separation techniques and cells carrying mutations in the CD95 palmitoylated site did not have an altered mobility in SDS-PAGE [28].
Membrane-enriched fractions contained readily detectable levels of ATP1A1 and also ICAM-2 proteins of the expected molecular mass using an antibody that recognizes the CD domain of ICAM-2 (sc-1512; Santa Cruz Biotech).
This was done for all the selected molecular gas masses using the present scan schedule as a trigger.
The sample (5 μL) was used for molecular mass determination using an ESI Q-TOF mass spectrometer (Micromass, Manchester, England).
The sample (5 μL) was used for molecular mass determination using JOEL MALDI-TOF.
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