Table 3.

Mastermix preparation for surface staining

Antigen	Metal	Clone	aDilution factor	Total [ul] amount for 330 μL / 11 sample	Total [ul] amount for 1,600 μL / 48 samples
CD3	115In	UCHT1	400	0.825	4
CD45	141Pr	HI30	400	0.825	4
CD19	142Nd	HIB19	400	0.825	4
CD123	143Nd	6H6	400	0.825	4
CD4	145Nd	RPA-T4	400	0.825	4
IgD	146Nd	IA6-2	200	1.65	8
CD20	147Sm	2H7	400	0.825	4
CD66a/c/e	149Sm	ASL-32	400	0.825	4
CD14	151Eu	M5E2	200	1.65	8
CD7	153Eu	CD7-6B7	100	3.3	16
CD1c	154Sm	L161	800	0.4125	2
CD45RA	155Gd	HI100	400	0.825	4
CD27	158Gd	L128	400	0.825	4
CD11c	159Tb	Bu15	400	0.825	4
CD38	167Er	HIT2	400	0.825	4
CD8	168Er	SK1	800	0.4125	2
CD24	169Tm	ML5	100	3.3	16
CD141	173Yb	1A4	400	0.825	4
HLA-DR	174Yb	L243	100	3.3	16
CD56	176Yb	N901	100	3.3	16
CD16	209Bi	3G8	100	3.3	16
CD41	89Y	HIP8	400	0.825	4
			Total volume	31.35	152
			Volume to aliquot + ∼20%	38	n/a

Antibody volumes needed to prepare surface stating cocktail for one batch (11 samples) and multibatch experiments, 4 batches (48 samples), taking into consideration a dilution factor for each antibody and final staining volume obtained in Table 2. In this case 35 μL of antibody mixture (with 20% excess) is aliquoted in one single tube. In total 4 tubes are prepared.

^aFinal antibody concentration depends on the cell type, antibody lot and conditions. Therefore, users should titrate the antibody according to their own setting.