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Since the molecular weights of LSU of Rubisco from P. yezoensis, P. haitanensis, B. fuscopurpurea and L. japonica have been reported to be equal, the total protein of P. yezoensis was used as the control in this study, to mark the position of Rubisco LSU.
It is thus evident that the flux for any given reaction within the translational pathway must equal the total protein synthesis flux, at least if translational reactions are assumed to be 100% efficient and do not occur in futile cycles.
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The initial glycan concentrations were all assumed to be zero, except the first initial 9 mannose glycan, whose concentration was set equal to the total protein concentration.
To ensure equal loading, the total protein concentration of each sample was determined by Bradford Protein Assay (Bio-Rad).
(B ) Quantitative analysis of the initial nuclear translocation rates (equal to the total protein translocation rate through NPCs) for the experiments described in (A ).
From the infected cells, bacteria were purified and equal amount of the total protein was loaded in a 15% SDS PAGE.
Equal aliquot of the total protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane.
Equal amounts of the total protein from each lysate were quantified using a bicinchoninic acid protein quantification kit (Sigma).
For Western blot analysis, equal amounts of the total protein content were separated by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE; 15%) and transferred onto membranes.
An equal amount of the total protein was subjected to electrophoresis and then transferred to nitrocellulose membranes, incubated in blocking buffer (1% bovine serum albumin in Tris-buffered saline with 0.1% Tween-20) for 2 hours.
Every cellulosome sample grown in 14N medium was mixed with the reference 15N-labeled cellulose cellulosome sample in equal proportions based on the total protein concentration.
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