Table 2.

Three-Day Assay Signal Window and Z-Factor Coefficient Determination for the IL-6–Induced pSTAT3-Y705 Activation High-Content Screening Assay

Class/Day	Plate	Max/Min	Mean	SD	CV	PL to PL	Z-factor	S:B ratio
Intraplate
1	1	Max	10,690.4	1,102.6	10.3	1–3	0.59	5.73
	2	Max	11,082.8	947.4	8.6	1–4	0.6	6.23
	3	Min	1,866.0	103.3	5.5	2–3	0.66	5.94
	4	Min	1,714.6	86.5	5.1	2–4	0.67	6.46
	All plates					All	0.61	6.08
2	1	Max	12,108.7	782.5	6.5	1–3	0.73	6.33
	2	Max	11,697.7	920.4	7.9	1–4	0.72	6.39
	3	Min	1,911.6	130.9	6.9	2–3	0.68	6.12
	4	Min	1,894.3	161.3	8.5	2–4	0.67	6.18
	All plates					All	0.69	6.26
3	1	Max	7,038.9	1,079.8	15.3	1–3	0.38	6.93
	2	Max	8,078.0	1,196.8	14.8	1–4	0.4	7.62
	3	Min	1,015.9	174.7	17.2	2–3	0.42	7.95
	4	Min	923.3	144.2	15.6	2–4	0.44	8.75
	All plates					All	0.35	7.8
Interplate
1	1 and 2	Max	10,886.6	1,046.3	9.6		0.61	6.08
	3 and 4	Min	1,790.3	121.7	6.8
2	1 and 2	Max	11,903.2	878.5	7.4		0.69	6.26
	3 and 4	Min	1,903.0	147.1	7.7
3	1 and 2	Max	7,558.4	1,252.5	16.6		0.35	7.80
	3 and 4	Min	969.6	166.7	17.2
Day to day
1 and 2	All plates	Max	11,394.9	1,091.5	9.6		0.61	6.17
		Min	1,846.7	146.2	7.9
2 and 3	All plates	Max	9,730.8	2,427.0	24.9		−0.06	6.78
		Min	1,436.3	492.5	34.3

 The average inner intensity pSTAT3-Y705 Ch 2 data presented in the table was generated in three independent experiments of 2×384-well plates each of the maximum (50 ng/mL IL-6 and 0.2% DMSO) and minimum (0.2% DMSO) plate controls conducted on three separate days.

 S:B, signal-to-background ratio.