Thereafter, the cells are washed once with staining buffer before being incubated in staining buffer supplemented with 5% FBS on ice until sorting is completed.
The resuspended cells were kept on ice until sorting.
Cells to be sorted were resuspended in complete medium and kept cold until sorting.
The staining was performed in batches during sorting until sorting was completed.
4,6-Diamidino-2-phenylindole (DAPI) was added (0.2 μg/mL) to the single-cell suspension, and cells were incubated on ice until sorting (about 30 minutes).
Following incubation, cells were centrifuged, and the cell pellets were resuspended in 0.5 ml ALDEFLOUR assay buffer and maintained on ice until sorting.
Cells were resuspended in 1X PBS, pH 7.4, supplemented with 2% bovine serum albumin (Sigma-Aldrich) and kept at 4°C until sorting.
The cell pellet was resuspended in 200 μL PBS/1%BSA with 10 μL phycoerythrin- (PE-) conjugated goat anti-mouse IgM antibody (R&D Systems, Minneapolis, MN) for 30 min on ice, washed for 3 times, resolved in 400 μL of PBS, and kept on ice until sorting.
Boycott United Airlines until they sort this out.
We put 30% in holding until we sort it out".
