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In the present study the influence of CH…O interactions in the conformational stability of penicillin binding proteins were analyzed in both Gram positive and Gram negative bacteria.
As a comparison with AcrB, two sterol binding proteins were analyzed.
To test this further, we carried out pulldown assays with biotinylated H2A, H2B, H3 and H4N-terminal peptides and HeLa nuclear extracts, and the resulting binding proteins were analyzed by western blot using antibody against Sin3A and CoREST.
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The biotin binding of the steroid-binding proteins was analyzed by the biotin-coated sensor chip.
Only cells expressing GAD and at least 1 gene encoding for 1 neuropeptide or 1 calcium-binding protein were analyzed.
For UHRF1 and FANCL in vitro DNA binding, the proteins were analyzed by electrophoresis on 10% SDS-PAGE gel followed by western blot analysis.
The spatial correlations between mA sites (RRACH) and ESEs or binding clusters of SR proteins were analyzed by BEDTools software.
Alignment analyses of the interested genes or proteins were performed by online software ClustalX (http://www.ebi.ac.uk/Tools/msa/clustalw2/), and functional domains and protein binding sites of interested proteins were analyzed by Conserved Domains Database (CDD) search program (http://www.ncbi.nlm.nih.gov/cdd).nih.gov/cdd
The precipitates were washed three times with binding buffer, and the bound proteins were analyzed by SDS PAGE and western blot with Ku80 as loading control.
The reaction was incubated overnight at 4°C, washed three times with binding buffer and the bound proteins were analyzed by 10% SDS-PAGE and autoradiography.
FcRn−, Protein A and CD16a-binding to recombinant IgG1-Fc proteins were analyzed by surface plasmon resonance (SPR) spectroscopy using a Biacore 3000 instrument (GE Healthcare, Waukesha, WI).
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