Protocol for immunophenotyping with directly conjugated mcabs

1. Aliquot equal numbers of cells from a cell pellet or 100 ml of whole blood into Falcon tubes.
2. Add the required directly conjugated mcabs (1 mg) to each aliquot of cells.
3. Mix on Vortex.
4. Incubate at RT for 20 min.
5. Wash cells in PBS if using isolated cells.
6. Resuspend cell pellet in <0.5 ml PBS.
7. If using whole blood add 1 ml of 1:10 diluted Becton Dickinson FACS Lysing Solution.
8. Mix on Vortex.
9. Incubate at RT for 10 min.
10. Collect least 10,000 events.