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In these studies, it is hypothesized that cancer cells which are autophagy-dependent are easier to treat, since removal of autophagy will remove the cell survival mechanisms.
After remove the cell debris with centrifuge, the crude EG enzyme solution was prepared.
With the purpose of improving the purity, surfactant was added to the standard condition to remove the cell materials further.
To remove the cell biomass, the resultant solution was centrifuged at 5000 rpm for 10 min (MiniSpin Eppendorf, USA).
The extracted coelomic fluid was pooled and centrifuged at 10,000 rpm for 15 min at 4 °C to remove the cell debris which settled down as pellet.
Decellularization by various detergents such as sodium dodecyl sulfate (SDS) and triton X-100 can remove the cell nuclei in tissue organs.
The successful production and purification of full-length, soluble, and natural fusion proteins, however, are still retarded by various obstacles such as the need for pretreatment to remove the cell debris and colloid contaminants, a relatively long operation time, and protein solubility.
Two types of chemicals were applied either solely or together to remove the cell materials: (1) alkalis (NaOH at concentrations varying between 0.02 M and 1 M, or 0.2 M NH4OH), and (2) surfactant (SDS at concentrations varying between 0.025% and 0.2%).
The lysates were incubated on ice for 20 min, sonicated and centrifuged to remove the cell debris.
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Briefly, SARS-CoV-infected 2B4 cells (MOI = 0.1) grown on the 24-well plates were washed with phosphate-buffered saline (PBS) at indicated time points after infection to remove the cell-free viruses.
To remove the cell-surface-associated HA, the cells were incubated for 10 min at 37 °C with trypsin-EDTA and washed with PBS.
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