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The eluted mixture was resolved by 10% SDS-PAGE and stained with Commassie Blue R250 and the bands were excised from the gel, followed by tryptic digestion and mass spectrometry.
The biotinylation mixture was resolved on 12% SDS-PAGE, transferred onto nitrocellulose membrane and detected by streptavidin HRP.
The digestion mixture was resolved from di- dp2 di- dp2a-(dp18) decasaccharide, and dp2 to dp8 were further purified by strocta- dp18exchange HPLC as described [37].
After boiling, the half of reaction mixture was resolved on a 10% denaturing gel and the radioactivity was detected by autoradiography.
Following extensive washes, the reaction mixture was resolved by SDS-PAGE electrophoresis and radioactive proteins were visualized by phosphoimaging (Instantimager, Packard).
The supernatant was concentrated at 13000 g, for 30 min at 20°C using 3kDa cut off spin filters (Millipore, Bellerica, MA, USA), the extracted protein mixture was resolved by 12.5% SDS-PAGE and the gel stained with Coomassie blue (BioRad).
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The reaction mixtures were resolved on a 7.5% SDS-polyacrylamide gel and immunoblotted with anti-ubiquitin antibody (Zymed).
After digestion, the reaction mixtures were resolved on the nonreducing tricine SDS-PAGE and stained with Coomassie Brilliant Blue.
Where indicated, fOS mixtures and fOS-AP mixtures were resolved by HPLC using an amine-bonded silica column (LiChrospher Amino 5 µm, 250 mm×4.6 mm, Sulpelco Inc).
After derivatisation with 2-aminopyridine (2-AP) as previously described [29], [34], oligosaccharide mixtures were resolved by HPLC using an amine-bonded silica column (LiChrospher Amino 5 µm, 250 mm ×4.6 mm, Sulpelco Inc).
For example, to evaluate the effect of NQTrp (Figure 1A) on the transformation of the Aβ into the toxic assemblies, the inhibitor was incubated with Aβ1 42 at increasing molar ratios, and the reaction mixtures were resolved on SDS-PAGE (Figure 1B).
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