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1. Remove cells from Valmark plates as usual. Wash x2 in FACS buffer: HBSS (no
Ca, Mg, no phenol red) with 1% BSA, pH 7.4.
2. Pellet cells in eppendorf microfuge tube and resuspend 106 cells in 25 μl FACS
buffer containing 2 mg/ml human IgG.
3. Add 25 μl of FACS buffer containing 10 μg/ml M1/70, 5C6, M1/18, or your
antibody. If you want the cells to be in Mn2+ buffer, use FACS buffer containing
200 μM Mn2+.
4. Incubate on ice for 30 min.
5. Wash x2 with FACS buffer
6. Resuspend 106 cells in 25 μl FACS buffer containing 2 mg/ml human IgG.