Inhibition of Neutrophil Collagenase/MMP-8 and Gelatinase B/MMP-9 and Protection against Endotoxin Shock

Figure 2

Establishment of the indirect ELISA for detection of plasma P2 concentrations. (a) Immunoreaction of purified antibody from two rabbits that were immunized with P2-KLH. LgT represents the logarithm of serum dilution factors. (b) Two-dimensional serial dilution method for selection of coating concentration of P2-BSA and reagent concentration of polyclonal anti-P2 antibody. Two columns of data exist under each antibody dilution condition. The white panels represent absorbance after binding of the antibody to the immobilized P2-BSA whereas the gray panels represent absorbance after binding of the antibody to immobilized P2-BSA in presence of 800 ng/mL P2 as a competitor. (c) Antibody specificity determination. (d) The standard curve with P2 concentrations ranging from 50 to 3200 ng/mL. B0 is OD value in the absence of P2 in plasma, B is OD value at serial concentrations of P2 in plasma, and C is P2 concentration. In this example, serial P2 concentrations were prepared in mouse plasma that had been diluted 30 times with prewarmed PBS at 80°C.

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