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Infected leaflets were incubated for 6 days as described above.
Inoculated leaflets were incubated with 12 h photoperiod.
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To determine the PS content in the outer leaflet, cells were incubated with 5 µl Annexin-V-FITC in 150 µl Ca2+ containing Annexin-V-binding Annexin-V-binding Annexin-V-bindingt room temperature in the dark.
To isolate plasma membrane vesicles, NIH3T3-tTG fibroblasts were incubated with 100 µg/ml biotin-labeled Concanavalin A (Sigma) to bind it to the outside leaflet of the plasma membrane.
To examine if the mode of induced cell death was apoptosis, macrophage monolayers were incubated with a range of supernatant volumes and assessed for detection of Phosphatidylserine exposure on the outer leaflet of the cell membrane by annexin V binding.
Cells were incubated for 48h.
Embryos were incubated until hatch.
Uninfected MDMs were incubated with FM 4-64FX, a formaldehyde/glutaraldehyde-fixable analog of FM 4 64, a lipophilic dye that fluoresces intensely on binding to the outer leaflet of lipid bilayers.
Inoculated leaflets were then incubated as described for the virulence tests.
We reasoned that if M5-DLO-containing liposomes are incubated with Con A, DLOs located in the outer leaflet of the vesicles will interact with the lectin whereas those in the inner leaflet will not.
They are incubated by it.
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CEO of Professional Science Editing for Scientists @ prosciediting.com