The second antibody is a PAB raised in a different species coupled to the enzyme of choice. This method, also called the unlabeled antibody method, uses a preformed soluble enzyme-antienzyme immune complex, which is named after the particular enzyme used. In this case, the SPAP method uses a soybean peroxidase-antiperoxidase complex. The conjugate was stored in the refrigerator at 4 o C without loss of activity. Idealized representation contrasting the immunoassays described in this report.
Figure represents wells of a microtiter plate coated with antigen.
Primary antibody in each case was L3, a MAB generated against lipoxygenase. 0.0 Figure 2. Enzymatic activity of soybean and horseradish peroxidases after exposure to 70 o C for one hour. This prediction was confirmed by comparative heat inactivation profiles of the two enzyme-antibody conjugates. Both the HRP and the SBP were coupled to their respective MABs, and heated for varying periods.