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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
  1. 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.