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Table 3 Kinetic parameters obtained from the optimization of the Peptidomatrix conditions with the peptides and antibodies specific to Na/K ATPase (see figure 4) (± SD, n = 3)

From: Competition between bound and free peptides in an ELISA-based procedure that assays peptides derived from protein digests

A: increasing attached peptide at two fixed levels of added antibody
  1 stAB Concentration 0.00033 serum/PBS 1 stAB Concentration 0.00066 serum/PBS  
Kd (μg/ml) 0.19 ± 0.01 0.1 ± 0.02  
Amax (OD units) 1.36 ± 0.02 1.56 ± 0.08  
D (OD units) 0.25 ± 0.02 0.38 ± 0.07  
B: Competition assay – Kd determined when increasing added free peptide at three fixed levels of added antibody
  1 stAB concentration 0.000625 serum/PBS 1 stAB concentration 0.000125serum/PBS 1 stAB concentration 0.0025 serum/PBS
Kd (μg/ml) 0.55 ± 0.1 0.83 ± 0.12 2.55 ± 1.21
Amax (OD units) 1.16 ± 0.05 1.35 ± 0.05 1.33 ± 0.17
D (OD units) 0.59 ± 0.04 0.86 ± 0.04 1.17 ± 0.18
C: increasing attached peptide at two fixed levels of added antibody
  1 stAB concentration 0.00033 serum/PBS 1 stAB concentration 0.00067 serum/PBS  
Kd (serum dilutions) 0.33 ± 0.08 0.29 ± 0.04  
Amax (OD units) 1.46 ± 0.09 1.74 ± 0.06  
D (OD units) 0.19 ± 0.079 0.24 ± 0.05  
D: Competition assay- Kd determined when increasing added free peptide at three fixed levels of added antibody
  1 stAB concentration 0.000625 serum/PBS 1 stAB concentration 0.00125 serum/PBS 1 stAB concentration 0.0025 serum/PBS
Kd (μg/ml) 0.68 ± 0.12 0.99 ± 0.19 2.04 ± 0.3
Amax (OD units) 1.58 ± 0.06 1.87 ± 0.09 2.05 ± 0.08
D (OD units) 0.25 ± 0.05 0.25 ± 0.08 0.28 ± 0.08
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Proteome Science

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