BMC Cancer

Table 1 Chessboard titrations of ELISA antibodies

From: Development of fragment-specific osteopontin antibodies and ELISA for quantification in human metastatic breast cancer

	2F10 μg/ml
fl-OPN	50	25	12.5	6.25	3.125	1.56	0.781	0.39	0.20	0.098	0.049	0
5	2.66	2.73	2.92	3.32	3.66	3.78	1.30	0.76	0.54	0.43	0.459	0.332
2.5	2.84	o.s.	3.07	o.s.	o.s.	3.89	1.16	0.60	0.40	0.29	0.276	0.18
1.25	2.62	o.s.	o.s.	3.44	o.s.	3.95	1.14	0.49	0.32	0.20	0.174	0.119
0.625	2.74	o.s.	o.s.	o.s.	o.s.	o.s.	1.17	0.34	0.24	0.17	0.154	0.086
0.313	2.86	o.s.	o.s.	o.s.	o.s.	o.s.	1.01	0.31	0.20	0.15	0.126	0.074
0.156	o.s.	o.s.	o.s.	o.s.	o.s.	o.s.	0.964	0.322	0.184	0.106	0.076	0.054
0.078	o.s.	o.s.	o.s.	o.s.	o.s.	o.s.	0.971	0.304	0.182	0.102	0.076	0.05
0	1.619	1.869	1.071	0.523	0.285	0.177	0.119	0.084	0.073	0.071	0.069	0.071
	1F11b μg/ml
fl-OPN	1.56	0.78	0.39	0.195	0.098	0.049	0.024	0.012	0.006	0.003	0.0015	0
0.156	3.66	3.80	3.801	3.77	3.74	3.61	2.16	1.473	0.77	0.41	0.254	0.041
0.078	3.83	o.s.	o.s.	3.93	o.s.	3.31	1.91	1.06	0.62	0.37	0.231	0.041
0.039	3.83	o.s.	o.s.	o.s.	3.96	3.07	2.36	1.047	0.56	0.32	0.185	0.035
0.020	3.84	o.s.	o.s.	o.s.	3.76	2.43	1.41	0.763	0.41	0.24	0.154	0.039
0.0098	3.86	o.s.	3.48	3.49	2.45	1.61	0.95	0.643	0.34	0.20	0.132	0.039
0.0049	3.82	3.81	2.79	2.27	1.53	0.96	0.50	0.358	0.22	0.16	0.142	0.036
0.002	3.05	2.40	1.54	1.18	0.90	0.60	0.32	0.215	0.13	0.087	0.07	0.04
0	0.46	0.26	0.17	0.10	0.069	0.05	0.05	0.04	0.03	0.04	0.038	0.038

Both the capture antibody (2F10, top) and the detecting antibody (1F11, bottom) were analyzed for dose response in a checkerboard analysis. The optimal concentrations (bold) are 1.56 μg/ml capture antibody and 0.098 μg/ml detecting antibody 1F11b. Conditions listed as o.s. (off-scale) had an absorbance > 4.0.

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