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The high antigen binding was due to a site-directed orientation of the thiol-biotinylated fragments.
To rule out that the DNA binding was due to the GFP-part of the fusion protein, the protein trap line G180 [12] that is localized to the nucleoplasm was permeabilized.
The addition of a YB-1 (1 µg) antibody caused a super-shift in the binding product, confirming that binding was due to this transcription factor (Figure 3F, lane 4).
Preincubation with mAb 7D2 significantly reduced HIV-1NL4-3 binding to IFN-α-treated monocytes comparable to that observed for non-induced monocytes, indicating that IFN-α-induced HIV-1NL4-3 binding was due to Sn alone.
We next determined if anti-Ig mediated downmodulation of gp41 MPER binding was due to internalization of gp41 MPER/Ig complexes by pre-incubating with anti-Ig Abs (Fig. 4E).
It could be possible that lack of binding was due to fast dissociation of the complex.
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Since this construct lacks the C-μ2 subdomain, it is unable to bind the YxxΦ motif and thus any observed binding is due to nonspecific interactions.
The reversible binding is due to molecular complementarity between the inhibitor and the active site of hAVCP, which confers the selectivity of the inhibitor.
In acetonitrile, blue shifts in fluorescent emission upon zinc binding are due to the formation of a 1 2 metal/ligand complex, which induced a fluorescent emission at 616 nm at the expense of the fluorescent emission at 672 nm.
Our results in Fig. 2d indicate that the K472E/R473E mutation per se does not inhibit CAND1 binding, but that decreased CAND1 binding is due to the increased neddylation levels.
To confirm that such binding is due to the recognition of U-runs by SNF, mutants of RNA D were constructed in which either or both U-runs were changed to UC-runs (Figure 2B).
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