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Cut pieces of membrane were washed in two changes of sterile (filtered and autoclaved) aged seawater with moderate shaking to remove unattached bacteria.
After the precipitation, the iron oxide nanoparticles (~10 nm) formed within the membrane were washed away by flowing DI water through the membrane using the vacuum filtration setup.
To analyze the amount of sorbed Fe(II), the solids on the membrane were washed with 3 ml Mill-Q water to remove sulfide residual on the solids, and then the solids along with the membrane were transferred into 3.0 ml of 0.5 M HCl solution and mixed with a magnetic Teflon bar for 20 min for Fe(II) desorption.
Migrating cells on the outside membrane were washed and stained with crystal violet for 10 min.
Membrane were washed twice with TBST for 10 min and incubated with secondary antibody for 1 h at room temperature.
The peptides on the membrane were washed twice with 200 µl of Tween-TBS (TTBS) and were processed similarly to a western blot method.
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The membrane was washed three times with PBST, and then visualized by excitation at 800 nm.
The membrane was washed with TBST for 5 min and incubated with a primary antibody.
Then the membrane was washed twice with phosphate buffered saline (PBS, pH 7.4, Sigma-Aldrich, Steinheim, Germany).
The membrane was washed again before visualizing with the Odyssey® classic infra-red imaging system (Li-Cor).
The membrane was washed to remove unbound material followed by incubation with a cocktail of biotinylated detection antibodies.
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CEO of Professional Science Editing for Scientists @ prosciediting.com